Protocol to screen chemicals to enhance homology-directed repair in CRISPR-Cas9 gene editing
Summary: CRISPR-Cas9-based gene editing via homology-directed repair (HDR) enables precise modifications, though its efficiency is limited by the prevalence of non-homologous end joining (NHEJ). Here, we present a protocol for enhancing HDR efficiency by identifying chemicals using high-throughput s...
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Elsevier
2025-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725003028 |
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| author | Min Ju Jo Hye Jin Nam |
| author_facet | Min Ju Jo Hye Jin Nam |
| author_sort | Min Ju Jo |
| collection | DOAJ |
| description | Summary: CRISPR-Cas9-based gene editing via homology-directed repair (HDR) enables precise modifications, though its efficiency is limited by the prevalence of non-homologous end joining (NHEJ). Here, we present a protocol for enhancing HDR efficiency by identifying chemicals using high-throughput screening (HTS). We describe steps for designing 96-well plates, executing HTS, and performing data analysis. We then detail procedures for identifying small molecules that improve HDR-associated gene editing. This protocol has potential application in HTS analysis focused on discovering reliable HDR enhancers.For complete details on the use and execution of this protocol, please refer to Jang et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-7a60cbd3855f4964aafb59e41ecc4fa1 |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-7a60cbd3855f4964aafb59e41ecc4fa12025-08-20T02:06:47ZengElsevierSTAR Protocols2666-16672025-09-016310389610.1016/j.xpro.2025.103896Protocol to screen chemicals to enhance homology-directed repair in CRISPR-Cas9 gene editingMin Ju Jo0Hye Jin Nam1Therapeutics & Biotechnology Division, Korea Research Institute of Chemical Technology, Daejeon 34114, Republic of Korea; Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon 34113, Republic of KoreaTherapeutics & Biotechnology Division, Korea Research Institute of Chemical Technology, Daejeon 34114, Republic of Korea; Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon 34113, Republic of Korea; Corresponding authorSummary: CRISPR-Cas9-based gene editing via homology-directed repair (HDR) enables precise modifications, though its efficiency is limited by the prevalence of non-homologous end joining (NHEJ). Here, we present a protocol for enhancing HDR efficiency by identifying chemicals using high-throughput screening (HTS). We describe steps for designing 96-well plates, executing HTS, and performing data analysis. We then detail procedures for identifying small molecules that improve HDR-associated gene editing. This protocol has potential application in HTS analysis focused on discovering reliable HDR enhancers.For complete details on the use and execution of this protocol, please refer to Jang et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725003028high-throughput screeningmolecular biologyCRISPR |
| spellingShingle | Min Ju Jo Hye Jin Nam Protocol to screen chemicals to enhance homology-directed repair in CRISPR-Cas9 gene editing STAR Protocols high-throughput screening molecular biology CRISPR |
| title | Protocol to screen chemicals to enhance homology-directed repair in CRISPR-Cas9 gene editing |
| title_full | Protocol to screen chemicals to enhance homology-directed repair in CRISPR-Cas9 gene editing |
| title_fullStr | Protocol to screen chemicals to enhance homology-directed repair in CRISPR-Cas9 gene editing |
| title_full_unstemmed | Protocol to screen chemicals to enhance homology-directed repair in CRISPR-Cas9 gene editing |
| title_short | Protocol to screen chemicals to enhance homology-directed repair in CRISPR-Cas9 gene editing |
| title_sort | protocol to screen chemicals to enhance homology directed repair in crispr cas9 gene editing |
| topic | high-throughput screening molecular biology CRISPR |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725003028 |
| work_keys_str_mv | AT minjujo protocoltoscreenchemicalstoenhancehomologydirectedrepairincrisprcas9geneediting AT hyejinnam protocoltoscreenchemicalstoenhancehomologydirectedrepairincrisprcas9geneediting |