Gene Editing

Technologies for Enhancing Plant Yield and Pest Resistance

1. Introduction: The Promise of Gene Editing for Garden Plants

The ability to precisely modify the genetic material of organisms has opened unprecedented avenues for enhancing desirable traits in plants. Gene editing technologies, with their capacity to alter the genomic architecture at specific locations with remarkable accuracy, hold significant potential for revolutionizing the characteristics of garden plants, particularly concerning crop yields and resistance to pests.¹ These advanced tools offer a pathway to address the escalating demands of a growing global population and to cultivate crops that can better withstand the challenges posed by changing environmental conditions, making them highly pertinent not only for large-scale agricultural endeavors but also for the more localized context of home gardens and small-scale farming.³ The development of these technologies signifies a notable shift towards more targeted and efficient strategies in plant breeding, moving beyond the inherent limitations of traditional methods.

2. Decoding Gene Editing Technologies

2.1. CRISPR-Cas9: The Revolution in Plant Modification

At the forefront of plant gene editing technologies stands the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) system. Derived from the natural defense mechanisms of bacteria against viral infections, CRISPR-Cas9 has rapidly become the dominant tool in the field due to its simplicity, efficiency, and versatility in making precise modifications to DNA sequences.² This system operates through the guidance of a small RNA molecule, known as the guide RNA (gRNA), which directs the Cas9 enzyme to a specific target DNA sequence within the plant's genome.⁶ Once the Cas9 enzyme reaches its designated site, it induces a double-stranded break (DSB) in the DNA molecule.⁶ The plant cell then employs its inherent DNA repair mechanisms, primarily Non-Homologous End Joining (NHEJ) and Homology Directed Repair (HDR), to fix these breaks.⁶ By manipulating these repair pathways, scientists can achieve various genetic modifications, including knocking out specific genes, inserting new genetic material, or making precise alterations to the existing DNA sequence.⁶ The RNA-guided nature of CRISPR-Cas9 offers a significant advantage in terms of design and adaptability compared to earlier protein-based systems, making it a highly versatile tool for plant genome engineering.

2.2. Beyond CRISPR: Exploring Other Gene Editing Tools

While CRISPR-Cas9 has garnered considerable attention, other gene editing technologies have also played crucial roles in advancing plant science and horticulture. Among these are Zinc Finger Nucleases (ZFNs) and Transcription Activator-Like Effector Nucleases (TALENs), which were developed prior to CRISPR and similarly function by inducing DSBs at specific DNA sequences.² ZFNs utilize custom-designed proteins that incorporate zinc finger domains to recognize and bind to specific DNA sequences, while TALENs employ Transcription Activator-Like Effector (TALE) domains for DNA targeting.¹⁴ Both ZFNs and TALENs are fused to a nuclease domain, typically from the FokI restriction enzyme, which cleaves the DNA at the targeted site.¹⁴ Although these technologies have been effective in plant genome editing, they are often more complex to design and may have lower efficiency compared to CRISPR-Cas9 in many applications.¹⁵

More recent advancements in gene editing include base editing and prime editing. Base editing allows for highly precise, targeted conversion of single nucleotide bases within the DNA sequence without inducing DSBs.² For example, adenine base editors (ABEs) can convert adenine-thymine (A-T) base pairs to guanine-cytosine (G-C) base pairs.² Similarly, cytosine base editors (CBEs) can facilitate the conversion of cytosine-guanine (C-G) to thymine-adenine (T-A).⁴⁸ Prime editing represents an even more versatile and precise approach, enabling various types of edits, including small insertions, deletions (indels), and all twelve possible base-to-base substitutions, often without requiring DSBs.¹⁰ These newer techniques offer advantages in terms of precision and potentially reduced off-target effects, expanding the toolkit for genome modification in plants.

3. Enhancing Productivity: Gene Editing for Increased Yield in Garden Plants

3.1. Mechanisms of Yield Improvement through Gene Editing

Gene editing technologies offer several avenues for enhancing the yield of garden plants by precisely modifying genes that govern various aspects of plant growth and development.²⁶ One key strategy involves manipulating genes related to photosynthetic efficiency, thereby increasing the plant's ability to convert sunlight into energy and biomass.¹⁸ Another approach focuses on genes involved in nutrient uptake and utilization, ensuring that plants can efficiently acquire the necessary resources for optimal growth and productivity.¹⁸ Modifications to genes controlling flowering time and plant architecture can also significantly impact yield. For instance, promoting earlier or more synchronized flowering can optimize the reproductive cycle, while altering plant structure, such as branching patterns or leaf orientation, can improve light capture and resource allocation.⁹ Furthermore, enhancing the plant's tolerance to various abiotic stresses, such as drought, salinity, and extreme temperatures, through gene editing can prevent substantial yield losses under unfavorable environmental conditions.¹ The precise nature of gene editing allows for targeted improvements in these complex traits, often by manipulating genes involved in crucial hormone signaling pathways, such as those regulated by abscisic acid, which plays a vital role in plant growth and stress responses.⁶⁵

3.2. Case Studies: Examples of Yield Enhancement in Specific Garden Plants

Several studies have demonstrated the successful application of gene editing to enhance the yield of plants. In rice, a staple crop globally, researchers utilized CRISPR-Cas9 technology to silence a suite of genes associated with the phytohormone abscisic acid, resulting in a significant 25-31% increase in grain yield in field tests.⁶⁵ Additionally, the mutation of the dense and erect panicle1 (DEP1) gene in Indica rice through CRISPR-Cas9 led to improvements in yield-related traits, including denser and more erect panicles and reduced plant height.⁹ Modifying genes that regulate stomatal density in rice has been shown to enhance drought tolerance without negatively affecting photosynthesis, which indirectly contributes to maintaining yield under water-scarce conditions.⁶⁴ In tomatoes, another popular garden plant, targeting the SELF-PRUNING5G (SP5G) gene using CRISPR-Cas9 resulted in rapid flowering and a more compact growth habit, leading to an earlier harvest.⁹ The application of CRISPR-Cas9 in maize involved introducing the native GOS2 promoter into the ARGOS8 gene, which enhanced grain yield under flowering stress conditions without any yield reduction in well-watered environments.⁶⁴ Researchers have employed CRISPR technology to modify genes responsible for leaf architecture and development in sugarcane, aiming to improve light capture efficiency and biomass production, which could potentially lead to higher sugar yields.⁶⁷ In soybean, inducing late flowering through CRISPR-Cas9 mediated mutation of the GmFT2a gene resulted in increased vegetative growth, which can often correlate with higher overall yields.⁹ These examples highlight the diverse ways in which gene editing can be employed to enhance the productivity of various garden plant species.

4. Fortifying Defenses: Gene Editing for Pest Resistance in Garden Plants

4.1. Strategies for Enhancing Pest Resistance at the Genetic Level

Gene editing offers a powerful and sustainable approach to fortify garden plants against a wide array of pests and diseases by precisely modifying their genetic makeup.²⁴ One primary strategy involves targeting host-susceptibility genes, which are genes that pathogens or pests exploit to infect or feed on the plant. By disrupting or modifying these genes using tools like CRISPR-Cas9, plants can become resistant to specific diseases or pests.²⁴ Another approach focuses on enhancing the plant's own resistance genes (R genes), which play a crucial role in the plant's immune system. Gene editing can be used to activate or modify these R genes to provide broader and more effective resistance against various pathogens.²⁴ Gene editing can be employed to enhance physical barriers to pest attacks, such as increasing the density or type of trichomes (small hairs) on plant surfaces, which can deter insect pests by affecting their feeding and movement.⁷⁰ In some cases, gene editing can even be used to develop plants that produce their own insecticidal proteins, offering a targeted and environmentally friendly way to control specific insect pests.²³ Researchers are exploring the use of gene editing to overcome insecticide resistance in pest populations, providing new avenues for integrated pest management strategies.²³ These multifaceted strategies underscore the potential of gene editing to create more resilient and pest-resistant garden plant varieties, reducing the reliance on chemical interventions.

4.2. Successful Applications: Examples of Pest-Resistant Gene-Edited Garden Plants

Several successful applications of gene editing have resulted in garden plants with enhanced resistance to common pests and diseases. Researchers have developed citrus plants that exhibit significant resistance to citrus canker, a serious bacterial disease, by using CRISPR-Cas9 to target the promoter region of the CsLOB1 gene, which is known to promote canker development.⁹ In grapevines, collaborative efforts are underway to utilize CRISPR technology to understand the genetic basis of resistance to powdery mildew, a prevalent fungal disease, with the aim of introducing this resistance into susceptible wine grape varieties.⁶⁷ Scientists are also working on using CRISPR to introduce beneficial mutations found in wild sorghum varieties into domesticated sorghum lines to enhance their resistance to various infestations, potentially offering a transgene-free solution for pest control.⁶⁷ A notable example in vegetable crops is the use of a cucumber CRISPR-Cas9 system to deactivate the eIF4E gene, which resulted in the creation of non-transgenic cucumber plants that showed resistance to multiple viruses, including Cucumber vein yellowing virus and potyviruses.¹¹ In rice, TALENs have been successfully employed to engineer resistance to bacterial blight, a destructive disease, by editing the regulatory region of the OsSWEET14 gene, which is essential for the pathogen to cause infection.⁷³ Similarly, wheat lines with enhanced resistance to powdery mildew have been developed using TALENs to target the TaMLO gene.³⁴ Genome editing has contributed to the production of banana mutants with the DMR6 gene, which confers resistance to banana wilt, a disease caused by Xanthomonas bacteria.⁶³ These successful examples highlight the broad applicability of gene editing in developing pest-resistant garden plant varieties.

5. Tangible Results: Successful Examples of Gene-Edited Garden Plants with Improved Traits

The application of gene editing technologies has yielded several garden plant varieties with notable improvements in yield and pest resistance.

5.1. Showcasing Plants with Enhanced Yield

• Rice: CRISPR-Cas9 mediated modifications in genes related to abscisic acid signaling resulted in a 25-31% increase in grain yield.⁶⁵ Mutation of the DEP1 gene in rice also improved yield-related traits.⁹

• Tomato: Editing the SP5G gene using CRISPR-Cas9 led to earlier flowering and a compact growth habit, resulting in an earlier yield.⁹

• Maize: Introduction of the native GOS2 promoter into the ARGOS8 gene via CRISPR-Cas9 enhanced grain yield under drought stress.⁶⁴

• Sugarcane: CRISPR-mediated modifications to genes responsible for leaf architecture improved light capture and biomass, potentially increasing sugar yields.⁶⁷

• Soybean: Mutation of the GmFT2a gene using CRISPR-Cas9 caused late flowering and increased vegetative size.⁹

5.2. Highlighting Plants with Robust Pest Resistance

• Citrus: CRISPR-Cas9 targeted modification of the CsLOB1 gene promoter conferred resistance to citrus canker.⁹

• Grapevine: Research using CRISPR is underway to introduce resistance to powdery mildew into susceptible varieties.⁶⁷

• Sorghum: CRISPR is being used to introduce wild-type resistance genes into domesticated varieties to enhance resistance to infestation.⁶⁷

• Cucumber: Deactivation of the eIF4E gene using CRISPR-Cas9 resulted in resistance to multiple viruses.¹¹

• Rice: TALEN-mediated editing of the OsSWEET14 gene engineered resistance to bacterial blight.⁷³

• Wheat: TALENs targeting the TaMLO gene developed lines with powdery mildew resistance.³⁴

• Banana: Genome editing of the DMR6 gene produced mutants resistant to banana wilt.⁶³

6. The Gardener's Perspective: Benefits and Drawbacks of Gene Editing for Home and Small-Scale Agriculture

6.1. Potential Advantages: Higher Yields, Reduced Pesticide Use, Improved Quality

For home gardeners and small-scale agriculturalists, gene editing technologies present a promising array of potential benefits. One of the most significant advantages is the possibility of achieving higher yields from their plants, allowing for greater food production within the same area.²⁶ This can be particularly valuable for those with limited gardening space. The enhanced resistance to pests and diseases conferred by gene editing could substantially reduce or even eliminate the need for chemical pesticides.⁴ This not only leads to healthier and more environmentally friendly gardening practices but also reduces the potential exposure to harmful chemicals for both the gardener and the consumers of the produce. Beyond yield and pest resistance, gene editing can also improve the overall quality of garden produce by enhancing nutritional content, flavor, texture, and extending shelf life.⁵ Compared to traditional plant breeding methods, which can take many years to achieve desired traits, gene editing offers a much faster and more precise route to developing improved plant varieties.³ This accelerated pace of development means that gardeners and small farmers could potentially have quicker access to plants with traits that better suit their needs and growing conditions.

6.2. Potential Disadvantages: Accessibility, Cost, Ethical Concerns, Regulatory Landscape

Despite the considerable potential benefits of gene editing for home gardeners and small-scale agriculture, several challenges and disadvantages currently exist. One of the primary limitations is the accessibility of these technologies. Currently, gene editing is largely confined to the realm of academic research institutions and large agricultural companies, making it difficult for the average gardener to directly utilize these tools.²⁵ The cost associated with developing and implementing gene editing in plants can also be substantial, which might translate to higher prices for gene-edited seeds or plants, potentially making them less affordable for home gardeners and small-scale farmers.⁶² Additionally, ethical concerns surrounding the genetic modification of plants and the potential long-term impacts on biodiversity may lead some gardeners to have reservations about using gene-edited varieties.⁴ The regulatory landscape for gene-edited plants is complex and varies significantly across different regions, which can create uncertainty and potentially hinder the availability of certain gene-edited varieties.⁴ While studies suggest that off-target effects from gene editing are minimal compared to naturally occurring mutations, concerns about unintended consequences still exist.⁶² The widespread adoption of a few high-performing gene-edited varieties could potentially lead to a reduction in overall genetic diversity within plant species, making them more vulnerable to future pests or diseases.⁶²

7. Navigating the Landscape: Regulations and Ethical Considerations for Gene-Edited Garden Plants

7.1. Global Regulatory Approaches to Gene-Edited Crops

The regulatory landscape governing gene-edited crops, including those relevant to garden plants, is characterized by significant heterogeneity across different regions of the world.⁴ Countries such as the United States, Canada, Argentina, Brazil, Chile, and Colombia have generally adopted a more permissive stance, often regulating gene-edited plants in a manner similar to conventionally bred varieties, particularly if the genetic modifications do not involve the introduction of foreign DNA.³ In the United States, the USDA's Animal and Plant Health Inspection Service (APHIS) exempts certain gene-edited plants from review based on their plant-pest risk, especially those with minor edits that could theoretically have been achieved through traditional breeding.⁷² In contrast, the European Union and New Zealand have implemented more stringent regulatory frameworks, typically classifying most gene-edited organisms as genetically modified organisms (GMOs) and subjecting them to extensive pre-market assessments and regulations.⁵ This distinction often hinges on whether the gene editing process introduces foreign DNA into the plant's genome, as traditional GMO regulations primarily target transgenic organisms.⁵ Other countries, like India, are still in the process of formulating their regulatory approaches to these emerging technologies.⁸¹ This lack of a globally harmonized regulatory framework presents challenges for international research collaboration, the trade of gene-edited agricultural products, and the widespread adoption of these innovations. The varying regulatory approaches reflect differing societal values and perspectives on the balance between the potential benefits and perceived risks associated with gene editing in agriculture.

7.2. Ethical Debates Surrounding Gene Editing in Agriculture

The application of gene editing technologies in agriculture has sparked a range of ethical debates that extend to the use of these tools in garden plants.⁴ One prominent concern revolves around the potential for unintended consequences and off-target effects, where the gene editing tools might inadvertently modify genes other than the intended targets, leading to unforeseen changes in the plant's characteristics or its interactions with the environment.²⁵ The long-term ecological impacts of releasing gene-edited plants into the environment are also a subject of ethical scrutiny, with questions raised about potential effects on biodiversity, gene flow to wild relatives, and the overall stability of ecosystems.⁴ Philosophical and religious perspectives often contribute to the debate, with some individuals expressing concerns about the "naturalness" of gene-edited organisms and whether humans have the right to alter the genetic code in this way.⁷⁶ Issues of social justice and equity are also central to the ethical discussion, including concerns about corporate control over the seed market, the potential for increased dependence of farmers on large agricultural companies, and the accessibility of these technologies to small-scale farmers and gardeners in developing countries.⁷⁵ The importance of transparency, clear labeling of gene-edited products, and ensuring consumers have the right to make informed choices are also frequently emphasized in ethical considerations.⁷² These multifaceted ethical debates play a significant role in shaping public perception, influencing regulatory decisions, and guiding the responsible development and application of gene editing in agriculture.

8. Delivering the Change: Methods for Introducing Gene Edits into Plants

8.1. Common Techniques: Agrobacterium-mediated Transformation, Biolistics

Introducing gene editing tools into plant cells is a critical step in the process of creating gene-edited garden plants. Two of the most commonly employed techniques for this purpose are Agrobacterium-mediated transformation and biolistics.⁵⁵ Agrobacterium-mediated transformation relies on the natural ability of the bacterium Agrobacterium tumefaciens to transfer a segment of its DNA, known as T-DNA, into the host plant's genome.⁶ Researchers insert the gene-editing machinery, such as the Cas9 gene and the guide RNA sequence, into the T-DNA region of a plasmid within Agrobacterium. When plant tissues or cells are exposed to these bacteria, the T-DNA, carrying the editing tools, is transferred and integrated into the plant's chromosomes, leading to the expression of the gene-editing components within the plant cells.⁶ This method is particularly effective for dicotyledonous plants and often results in stable integration of the editing construct into the plant's genome.⁶⁰

Biolistics, also known as particle bombardment or the gene gun method, involves physically delivering gene-editing tools into plant cells using high-velocity microparticles.⁶ In this technique, DNA, RNA, or protein components of the gene-editing system are coated onto tiny gold or tungsten particles. These coated particles are then propelled at high speed towards plant tissues or cells, penetrating the cell walls and membranes and delivering the gene-editing tools into the cytoplasm and nucleus.⁶ Biolistics can be applied to a broader range of plant species, including monocotyledonous plants, which are often less amenable to Agrobacterium-mediated transformation.²¹ It also offers the flexibility to deliver various forms of gene-editing reagents, including DNA-free options like pre-assembled ribonucleoprotein complexes (RNPs).⁵⁶

8.2. Considerations for Practical Application

While Agrobacterium-mediated transformation and biolistics are the most common methods for delivering gene editing tools to plants, other techniques exist, including protoplast transfection, microinjection, viral vectors, and nanoparticles.²¹ Protoplast transfection involves delivering DNA or RNA into plant cells that have had their cell walls removed, often using chemical agents like polyethylene glycol (PEG) or physical methods like electroporation.⁸⁸ Microinjection allows for the direct injection of gene-editing reagents into individual plant cells, typically used for embryos or specialized tissues.⁸⁸ Viral vectors can be engineered to carry and deliver gene-editing components into plant cells through viral infection.⁵⁷ Nanoparticles are also being explored as potential delivery vehicles for gene-editing tools, offering the possibility of tissue-culture-free delivery in some cases.⁵⁶

For home gardeners, the direct application of these gene editing delivery methods is currently not feasible due to the requirement for specialized laboratory equipment, sterile conditions, and technical expertise.²¹ The primary impact of these technologies on home gardening will likely be through the availability of improved seeds or plants that have already undergone gene editing by researchers and agricultural companies. The focus for home gardeners will be on accessing these advanced varieties rather than performing the gene editing process themselves.

9. Conclusion: The Future of Gene-Edited Garden Plants

Gene editing technologies, with CRISPR-Cas9 at the forefront, hold transformative potential for enhancing plants by improving key traits such as yield and pest resistance, alongside other desirable characteristics like nutritional content and shelf life.⁴ Ongoing research and development are continuously refining these tools, leading to increased precision, efficiency, and a broader range of editing capabilities, as seen in advancements like base editing and prime editing.¹⁰ Addressing the existing ethical and regulatory challenges will be paramount to ensuring the responsible and widespread adoption of gene-edited garden plants.⁴ For home gardeners and small-scale agriculture, the future likely holds the promise of greater access to a diverse selection of gene-edited seeds and plants with traits tailored to enhance productivity, promote sustainable practices through reduced pesticide use, and improve the nutritional value of homegrown produce.²¹

Information for this article came from the Internet. As is well know some information from the Internet is suspect, although our intent was to produce an accurate article, understand some information may be based on someones opinion and may not be factual.

Works used

1. pmc.ncbi.nlm.nih.gov, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC5748717/#:~:text=Genome%20editing%20tools%20have%20the,to%20biotic%20and%20abiotic%20stresses.

2. Genome Editing Tools in Plants - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC5748717/

3. Genome Editing in Agricultural Biotechnology - FDA, accessed April 13, 2025, https://www.fda.gov/food/agricultural-biotechnology/genome-editing-agricultural-biotechnology

4. Plant breeding advancements with “CRISPR-Cas” genome editing technologies will assist future food security - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2023.1133036/full

5. Gene Editing Technologies In Plant Science: Between Controversy & Promises, accessed April 13, 2025, https://igrownews.com/gene-editing-technologies-in-plant-science-between-controversy-promises/

6. Genome Editing in Plants with CRISPR/Cas9 - Sigma-Aldrich, accessed April 13, 2025, https://www.sigmaaldrich.com/US/en/technical-documents/protocol/genomics/advanced-gene-editing/genome-editing-in-plants-with-crispr-cas9

7. What are genome editing and CRISPR-Cas9?: MedlinePlus Genetics, accessed April 13, 2025, https://medlineplus.gov/genetics/understanding/genomicresearch/genomeediting/

8. Principles, Applications, and Biosafety of Plant Genome Editing Using CRISPR-Cas9, accessed April 13, 2025, https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2020.00056/full

9. Plant Breeding Innovation: CRISPR-Cas9 | ISAAA.org, accessed April 13, 2025, https://www.isaaa.org/resources/publications/pocketk/54/

10. Advances in CRISPR/Cas technologies and their application in plants, accessed April 13, 2025, https://www.maxapress.com/article/id/63dcaabbfa6c583b06e7b1ee

11. CRISPR/Cas-Mediated Genome Engineering in Plants: Application and Prospectives - MDPI, accessed April 13, 2025, https://www.mdpi.com/2223-7747/13/14/1884

12. Versatile Applications of the CRISPR/Cas Toolkit in Plant Pathology and Disease Management - APS Journals, accessed April 13, 2025, https://apsjournals.apsnet.org/doi/10.1094/PHYTO-08-20-0322-IA

13. Gene Editing and Crop Improvement Using CRISPR-Cas9 System - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2017.01932/full

14. Full article: Genome Editing—Principles and Applications for Functional Genomics Research and Crop Improvement - Taylor & Francis Online, accessed April 13, 2025, https://www.tandfonline.com/doi/full/10.1080/07352689.2017.1402989

15. A Revolution toward Gene-Editing Technology and Its Application to Crop Improvement, accessed April 13, 2025, https://www.mdpi.com/1422-0067/21/16/5665

16. Enhancing Horticultural Crops through Genome Editing | Encyclopedia MDPI, accessed April 13, 2025, https://encyclopedia.pub/entry/47815

17. Gene editing in plants: progress and challenges - Oxford Academic, accessed April 13, 2025, https://academic.oup.com/nsr/article/6/3/421/5290356

18. Recent advances of CRISPR-based genome editing for enhancing staple crops - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC11456538/

19. Recent advances of CRISPR-based genome editing for enhancing staple crops - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2024.1478398/full

20. Genome editing for vegetable crop improvement: Challenges and future prospects, accessed April 13, 2025, https://www.frontiersin.org/journals/genetics/articles/10.3389/fgene.2022.1037091/full

21. CRISPR in Agriculture - Innovative Genomics Institute (IGI), accessed April 13, 2025, https://innovativegenomics.org/crisprpedia/crispr-in-agriculture/

22. Gene Editing for Plant Resistance to Abiotic Factors: A Systematic Review - MDPI, accessed April 13, 2025, https://www.mdpi.com/2223-7747/12/2/305

23. CRISPR-Cas Genome Editing for Insect Pest Stress Management in Crop Plants - MDPI, accessed April 13, 2025, https://www.mdpi.com/2673-7140/2/4/34

24. CRISPR gene editing to improve crop resistance to parasitic plants - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/genome-editing/articles/10.3389/fgeed.2023.1289416/full

25. Gene editing in plants: progress and challenges - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC8291443/

26. Applications, Benefits, and Challenges of Genome Edited Crops - The Council for Agricultural Science and Technology, accessed April 13, 2025, https://cast-science.org/wp-content/uploads/2024/03/CAST_IP74-Gene-Edited-Crops.pdf

27. Genetic Engineering and Genome Editing Advances to Enhance Floral Attributes in Ornamental Plants: An Update - PubMed Central, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC10707928/

28. Genome Editing in Plants: Exploration of Technological Advancements and Challenges, accessed April 13, 2025, https://www.mdpi.com/2073-4409/8/11/1386

29. Application of CRISPR-Cas9 genome editing technology in various fields: A review - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC10916045/

30. Genome editing for grass improvement and future agriculture ..., accessed April 13, 2025, https://academic.oup.com/hr/article/12/2/uhae293/7822275

31. Advancements in Plant Gene Editing Technology: From Construct Design to Enhanced Transformation Efficiency - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC11653234/

32. Enhancing Horticultural Crops through Genome Editing: Applications, Benefits, and Considerations - MDPI, accessed April 13, 2025, https://www.mdpi.com/2311-7524/9/8/884

33. A Critical Review: Recent Advancements in the Use of CRISPR/Cas9 Technology to Enhance Crops and Alleviate Global Food Crises - PubMed Central, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC8929161/

34. A Revolution toward Gene-Editing Technology and Its Application to Crop Improvement, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC7461041/

35. Recent Advances in Tomato Gene Editing - MDPI, accessed April 13, 2025, https://www.mdpi.com/1422-0067/25/5/2606

36. Application of ZFNs and TALENs for Improvement in Horticultural Crops - ResearchGate, accessed April 13, 2025, https://www.researchgate.net/publication/372419101_Application_of_ZFNs_and_TALENs_for_Improvement_in_Horticultural_Crops

37. Chapter -1 Application of ZFNS and Talens for Improvement in Cereal Crops, accessed April 13, 2025, https://www.researchgate.net/publication/373632632_Chapter_-1_Application_of_ZFNS_and_Talens_for_Improvement_in_Cereal_Crops

38. Applications and Major Achievements of Genome Editing in Vegetable Crops: A Review, accessed April 13, 2025, https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2021.688980/full

39. Progress in gene editing tools, implications and success in plants: a review - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/genome-editing/articles/10.3389/fgeed.2023.1272678/full

40. ZFN, TALEN and CRISPR/Cas-based methods for genome engineering - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC3694601/

41. What is Genome Editing: Techniques & Applications | Synthego, accessed April 13, 2025, https://www.synthego.com/learn/genome-editing-engineering

42. Advancing crop disease resistance through genome editing: a promising approach for enhancing agricultural production - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/genome-editing/articles/10.3389/fgeed.2024.1399051/full

43. CRISPR-Cas9, TALENs and ZFNs - the battle in gene editing | Proteintech Group, accessed April 13, 2025, https://www.ptglab.com/news/blog/crispr-cas9-talens-and-zfns-the-battle-in-gene-editing/

44. Expanding the genetic editing tool kit: ZFNs, TALENs, and CRISPR-Cas9 - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC4191047/

45. A schematic outline for generation of ZFN, TALEN, and CRISPR/Cas9... - ResearchGate, accessed April 13, 2025, https://www.researchgate.net/figure/A-schematic-outline-for-generation-of-ZFN-TALEN-and-CRISPR-Cas9-mediated-genome-edited_fig6_299419009

46. Comparison between ZFNs, TALENs, and CRRISR/Cas systems for genome editing, accessed April 13, 2025, https://www.researchgate.net/figure/Comparison-between-ZFNs-TALENs-and-CRRISR-Cas-systems-for-genome-editing_tbl2_277353736

47. pubmed.ncbi.nlm.nih.gov, accessed April 13, 2025, https://pubmed.ncbi.nlm.nih.gov/36479615/#:~:text=Base%20editing%20and%20prime%20editing%20are%20two%20newly%20developed%20precision,and%20donor%20repair%20templates%2C%20respectively.

48. Back to Basics - Base & Prime Editing - Front Line Genomics, accessed April 13, 2025, https://frontlinegenomics.com/back-to-basics-base-and-prime-editing/

49. Base Editing in Plants: Applications, Challenges, and Future Prospects - ResearchGate, accessed April 13, 2025, https://www.researchgate.net/publication/353482712_Base_Editing_in_Plants_Applications_Challenges_and_Future_Prospects

50. Applications of base editing in plants. | Download Scientific Diagram - ResearchGate, accessed April 13, 2025, https://www.researchgate.net/figure/Applications-of-base-editing-in-plants_tbl1_353482712

51. Prime Editing Technology and Its Prospects for Future Applications in Plant Biology Research, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC10530660/

52. Base Editing in Plants: Applications, Challenges, and Future Prospects - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2021.664997/full

53. Applications and Prospects of CRISPR/Cas9-Mediated Base Editing in Plant Breeding, accessed April 13, 2025, https://www.mdpi.com/1467-3045/45/2/59

54. Precise plant genome editing using base editors and prime editors - PubMed, accessed April 13, 2025, https://pubmed.ncbi.nlm.nih.gov/34518669/

55. Advances in Delivery Mechanisms of CRISPR Gene-Editing Reagents in Plants - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/genome-editing/articles/10.3389/fgeed.2022.830178/full

56. Strategies for delivery of CRISPR/Cas-mediated genome editing to obtain edited plants directly without transgene integration - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/genome-editing/articles/10.3389/fgeed.2023.1209586/full

57. Attaining the promise of plant gene editing at scale - PNAS, accessed April 13, 2025, https://www.pnas.org/doi/10.1073/pnas.2004846117

58. News: Revolving the Gene-Editing Landscape with Circular RNA-mediated Prime Editors, accessed April 13, 2025, https://crisprmedicinenews.com/news/revolving-the-gene-editing-landscape-with-circular-rna-mediated-prime-editors/

59. Optimization of Prime Editing in Rice, Peanut, Chickpea, and Cowpea Protoplasts by Restoration of GFP Activity - MDPI, accessed April 13, 2025, https://www.mdpi.com/1422-0067/23/17/9809

60. Advances in Delivery Mechanisms of CRISPR Gene-Editing Reagents in Plants - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC8819002/

61. Genome Editing: A Promising Path Toward More Sustainable Agriculture, accessed April 13, 2025, https://www.syngentavegetables.com/news/innovation-impact/genome-editing-promising-path-toward-more-sustainable-agriculture

62. CRISPR in agriculture: applications, benefits & risks - Automata, accessed April 13, 2025, https://automata.tech/blog/crispr-agriculture/

63. CRISPR-Cas Genome Editing for Horticultural Crops Improvement: Advantages and Prospects - MDPI, accessed April 13, 2025, https://www.mdpi.com/2311-7524/9/1/38

64. Genome editing techniques in plants: a comprehensive review and future prospects toward zero hunger, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC9208631/

65. CRISPR-edited rice plants produce major boost in grain yield - Purdue University News, accessed April 13, 2025, https://www.purdue.edu/newsroom/archive/releases/2018/Q2/crispr-edited-rice-plants-produce-major-boost-in-grain-yield.html

66. Gene Editing and its Applications | Plant Physiology - Oxford Academic, accessed April 13, 2025, https://academic.oup.com/plphys/pages/gene-editing-and-its-applications-focus-collections

67. CRISPR in Agriculture: 2024 in Review - Innovative Genomics Institute (IGI), accessed April 13, 2025, https://innovativegenomics.org/news/crispr-in-agriculture-2024/

68. Genome editing for plant disease resistance: applications and perspectives | Philosophical Transactions of the Royal Society B: Biological Sciences - Journals, accessed April 13, 2025, https://royalsocietypublishing.org/doi/10.1098/rstb.2018.0322

69. Genome editing for plant disease resistance: applications and perspectives | Philosophical Transactions of the Royal Society B: Biological Sciences - Journals, accessed April 13, 2025, https://royalsocietypublishing.org/doi/abs/10.1098/rstb.2018.0322

70. Molecular mechanisms, genetic mapping, and genome editing for insect pest resistance in field crops - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC9729161/

71. Unveiling the Genetic Symphony: Harnessing CRISPR-Cas Genome Editing for Effective Insect Pest Management - PubMed Central, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC10708123/

72. United States: Crops / Food - Global Gene Editing Regulation Tracker, accessed April 13, 2025, https://crispr-gene-editing-regs-tracker.geneticliteracyproject.org/united-states-crops-food/

73. Pocket K No. 59: Plant Breeding Innovation: TALENs - ISAAA.org, accessed April 13, 2025, https://www.isaaa.org/resources/publications/pocketk/59/default.asp

74. CRISPR Variants for Gene Editing in Plants: Biosafety Risks and Future Directions - MDPI, accessed April 13, 2025, https://www.mdpi.com/1422-0067/24/22/16241

75. Genome Editing - PSB VIB-UGent, accessed April 13, 2025, https://www.psb.ugent.be/research/genome-editing

76. Ethical Considerations in Genetic Engineering | Plantae, accessed April 13, 2025, https://plantae.org/ethical-considerations-in-genetic-engineering/

77. Ethical and legal implications of gene editing in plant breeding: a systematic literature review - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC10710910/

78. Ethical and legal implications of gene editing in plant breeding: a systematic literature review - Universiti Malaya, accessed April 13, 2025, https://dialogue.um.edu.my/WoS/30.pdf

79. What are the Ethical Concerns of Genome Editing?, accessed April 13, 2025, https://www.genome.gov/about-genomics/policy-issues/Genome-Editing/ethical-concerns

80. Gene-Edited Plants: Regulation and Issues for Congress, accessed April 13, 2025, https://crsreports.congress.gov/product/pdf/R/R47683/2

81. The evolving landscape around genome editing in agriculture: Many countries have exempted or move to exempt forms of genome editing from GMO regulation of crop plants, accessed April 13, 2025, https://www.embopress.org/doi/10.15252/embr.202050680

82. Gene-Edited Plants: Regulation and Issues for Congress, accessed April 13, 2025, https://sgp.fas.org/crs/misc/R47683.pdf

83. What are the risks of using CRISPR in plants? - YouTube, accessed April 13, 2025, https://www.youtube.com/watch?v=qlqUaVcstu8

84. Strategies for delivery of CRISPR/Cas-mediated genome editing to obtain edited plants directly without transgene integration - PubMed Central, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC10398581/

85. www.frontiersin.org, accessed April 13, 2025, https://www.frontiersin.org/journals/genome-editing/articles/10.3389/fgeed.2023.1209586/full#:~:text=The%20primary%20tools%20to%20deliver,degrees%20based%20on%20the%20crop

86. Plant biomacromolecule delivery methods in the 21st century - Frontiers, accessed April 13, 2025, https://www.frontiersin.org/journals/genome-editing/articles/10.3389/fgeed.2022.1011934/full

87. Strategic transgene-free approaches of CRISPR-based genome editing in plants - PMC, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC9958309/

88. DNA-free gene editing in plants: a brief overview - Taylor and Francis, accessed April 13, 2025, https://www.tandfonline.com/doi/full/10.1080/13102818.2020.1858159

89. pmc.ncbi.nlm.nih.gov, accessed April 13, 2025, https://pmc.ncbi.nlm.nih.gov/articles/PMC8819002/#:~:text=Polyethylene%20glycol%20(PEG)%2Dmediated,culture%20procedures%20(Figure%202C).

90. CRISPR Delivery Methods | IDT - Integrated DNA Technologies, accessed April 13, 2025, https://www.idtdna.com/pages/technology/crispr/crispr-delivery

91. CRISPR/Cas Delivery Methods in Plants | 5 - Taylor & Francis eBooks, accessed April 13, 2025, https://www.taylorfrancis.com/chapters/edit/10.1201/9781003247685-5/crispr-cas-delivery-methods-plants-neetu-singh-kushwah-shristi-verma-meenal-rathore-np-singh

92. Plant Genetic Engineering kit I plan to use to modify houseplants. : r/DIYbio - Reddit, accessed April 13, 2025, https://www.reddit.com/r/DIYbio/comments/10o8l3f/plant_genetic_engineering_kit_i_plan_to_use_to/