Enhancing Specificity, Precision, Accessibility, Flexibility, and Safety to Overcome Traditional CRISPR/Cas Editing Challenges and Shape Future Innovations
Abstract Derived from the bacterial immune system, CRISPR/Cas9 induces DSBs at specific DNA sequences, which are repaired by the cell's endogenous mechanisms, leading to gene insertions, deletions, or substitutions. Despite its transformative potential, several challenges remain in optimizing o...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2025-07-01
|
| Series: | Advanced Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/advs.202416331 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1839617842208571392 |
|---|---|
| author | Muna Alariqi Mohamed Ramadan Lu Yu Fengjiao Hui Amjad Hussain Xiaofeng Zhou Yu Yu Xianlong Zhang Shuangxia Jin |
| author_facet | Muna Alariqi Mohamed Ramadan Lu Yu Fengjiao Hui Amjad Hussain Xiaofeng Zhou Yu Yu Xianlong Zhang Shuangxia Jin |
| author_sort | Muna Alariqi |
| collection | DOAJ |
| description | Abstract Derived from the bacterial immune system, CRISPR/Cas9 induces DSBs at specific DNA sequences, which are repaired by the cell's endogenous mechanisms, leading to gene insertions, deletions, or substitutions. Despite its transformative potential, several challenges remain in optimizing of CRISPR/Cas systems, including off‐target effects, delivery methods, PAM restrictions, and the limitations of traditional editing approaches. This review focuses on the interplay between these challenges and their contributions to gene editing precision, specificity, accessibility, flexibility, and safety. How reducing off‐target effects enhances specificity and safety is explored, while discussing the role of HDR‐based editing in achieving precise gene modifications, alongside alternative methods such as base editing and prime editing. Improved delivery mechanisms are examined for their impact on accessibility and efficiency, while the reduction of PAM restrictions is highlighted for its contributions to flexibility. Lastly, emerging cleavage‐free editing technologies are evaluated as they relate to safety and accessibility. This focused review aims to clarify the connections among these aspects and outline future research directions for advancing CRISPR‐based applications. |
| format | Article |
| id | doaj-art-4b64b6ee7b094b38b7330cf8e0510f30 |
| institution | Matheson Library |
| issn | 2198-3844 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj-art-4b64b6ee7b094b38b7330cf8e0510f302025-07-24T12:06:18ZengWileyAdvanced Science2198-38442025-07-011228n/an/a10.1002/advs.202416331Enhancing Specificity, Precision, Accessibility, Flexibility, and Safety to Overcome Traditional CRISPR/Cas Editing Challenges and Shape Future InnovationsMuna Alariqi0Mohamed Ramadan1Lu Yu2Fengjiao Hui3Amjad Hussain4Xiaofeng Zhou5Yu Yu6Xianlong Zhang7Shuangxia Jin8Hubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan 430070 ChinaDepartment of Plant Genetic Resources Division of Ecology and Dry Land Agriculture Desert Research Center Cairo EgyptHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan 430070 ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan 430070 ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan 430070 ChinaCotton Research Institute Xinjiang Academy of Agriculture and Reclamation Science Shihezi Xinjiang 832000 ChinaCotton Research Institute Xinjiang Academy of Agriculture and Reclamation Science Shihezi Xinjiang 832000 ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan 430070 ChinaHubei Hongshan Laboratory National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan 430070 ChinaAbstract Derived from the bacterial immune system, CRISPR/Cas9 induces DSBs at specific DNA sequences, which are repaired by the cell's endogenous mechanisms, leading to gene insertions, deletions, or substitutions. Despite its transformative potential, several challenges remain in optimizing of CRISPR/Cas systems, including off‐target effects, delivery methods, PAM restrictions, and the limitations of traditional editing approaches. This review focuses on the interplay between these challenges and their contributions to gene editing precision, specificity, accessibility, flexibility, and safety. How reducing off‐target effects enhances specificity and safety is explored, while discussing the role of HDR‐based editing in achieving precise gene modifications, alongside alternative methods such as base editing and prime editing. Improved delivery mechanisms are examined for their impact on accessibility and efficiency, while the reduction of PAM restrictions is highlighted for its contributions to flexibility. Lastly, emerging cleavage‐free editing technologies are evaluated as they relate to safety and accessibility. This focused review aims to clarify the connections among these aspects and outline future research directions for advancing CRISPR‐based applications.https://doi.org/10.1002/advs.202416331CRISPR/Casdelivery systemsdouble‐stranded breaksgenome editingHDRoff‐target |
| spellingShingle | Muna Alariqi Mohamed Ramadan Lu Yu Fengjiao Hui Amjad Hussain Xiaofeng Zhou Yu Yu Xianlong Zhang Shuangxia Jin Enhancing Specificity, Precision, Accessibility, Flexibility, and Safety to Overcome Traditional CRISPR/Cas Editing Challenges and Shape Future Innovations Advanced Science CRISPR/Cas delivery systems double‐stranded breaks genome editing HDR off‐target |
| title | Enhancing Specificity, Precision, Accessibility, Flexibility, and Safety to Overcome Traditional CRISPR/Cas Editing Challenges and Shape Future Innovations |
| title_full | Enhancing Specificity, Precision, Accessibility, Flexibility, and Safety to Overcome Traditional CRISPR/Cas Editing Challenges and Shape Future Innovations |
| title_fullStr | Enhancing Specificity, Precision, Accessibility, Flexibility, and Safety to Overcome Traditional CRISPR/Cas Editing Challenges and Shape Future Innovations |
| title_full_unstemmed | Enhancing Specificity, Precision, Accessibility, Flexibility, and Safety to Overcome Traditional CRISPR/Cas Editing Challenges and Shape Future Innovations |
| title_short | Enhancing Specificity, Precision, Accessibility, Flexibility, and Safety to Overcome Traditional CRISPR/Cas Editing Challenges and Shape Future Innovations |
| title_sort | enhancing specificity precision accessibility flexibility and safety to overcome traditional crispr cas editing challenges and shape future innovations |
| topic | CRISPR/Cas delivery systems double‐stranded breaks genome editing HDR off‐target |
| url | https://doi.org/10.1002/advs.202416331 |
| work_keys_str_mv | AT munaalariqi enhancingspecificityprecisionaccessibilityflexibilityandsafetytoovercometraditionalcrisprcaseditingchallengesandshapefutureinnovations AT mohamedramadan enhancingspecificityprecisionaccessibilityflexibilityandsafetytoovercometraditionalcrisprcaseditingchallengesandshapefutureinnovations AT luyu enhancingspecificityprecisionaccessibilityflexibilityandsafetytoovercometraditionalcrisprcaseditingchallengesandshapefutureinnovations AT fengjiaohui enhancingspecificityprecisionaccessibilityflexibilityandsafetytoovercometraditionalcrisprcaseditingchallengesandshapefutureinnovations AT amjadhussain enhancingspecificityprecisionaccessibilityflexibilityandsafetytoovercometraditionalcrisprcaseditingchallengesandshapefutureinnovations AT xiaofengzhou enhancingspecificityprecisionaccessibilityflexibilityandsafetytoovercometraditionalcrisprcaseditingchallengesandshapefutureinnovations AT yuyu enhancingspecificityprecisionaccessibilityflexibilityandsafetytoovercometraditionalcrisprcaseditingchallengesandshapefutureinnovations AT xianlongzhang enhancingspecificityprecisionaccessibilityflexibilityandsafetytoovercometraditionalcrisprcaseditingchallengesandshapefutureinnovations AT shuangxiajin enhancingspecificityprecisionaccessibilityflexibilityandsafetytoovercometraditionalcrisprcaseditingchallengesandshapefutureinnovations |