Reference - Detail
| RRC ID | 87613 |
|---|---|
| Author | Marsic T, Gundra SR, Aouida M, Masood M, Salibi A, Schmidt F, Alquwayzani R, Mahfouz MM. |
| Title | Precise, specific gene editing via a compact GoCas12m-FokI chimeric nuclease. |
| Journal | Nucleic Acids Res |
| Abstract |
CRISPR gene editing technologies have transformed functional genomics and biotechnology. Despite these advances, challenges such as limited delivery capacity and off-target activity continue to hinder their therapeutic translation. We developed a chimeric gene editing platform by fusing the compact, catalytically inactive Cas12m guiding module (GoCas12m) with the FokI nuclease domain. GoCas12m-FokI system integrates the programmable DNA-binding capability of Cas12m with the dimerization-dependent cleavage mechanism of FokI, enabling precise genome editing. Our engineered XTEN-fused GoCas12m-FokI editor exhibits robust activity on both surrogate reporters and endogenous human loci, achieving high-efficiency editing at clinically relevant targets-including CLTA1, HBB, AIFM1, and ABL with no detectable off-target activity at in silico-predicted sites, as confirmed by targeted deep sequencing. Notably, GoCas12m-FokI is nearly half the size of conventional Cas9- or Cas12a-based editors, facilitating delivery via adeno-associated virus and other cargo-limited vectors. This dual-guided editor showed comparable editing efficiency to previously reported FokI-dCas9 systems on endogenous loci, while possessing a different PAM requirement and domain orientation. By combining compact architecture, high specificity, and modular programmability, the GoCas12m-FokI editor offers a powerful alternative for therapeutic genome editing and a promising tool for in vivo gene therapy applications. |
| Volume | 54(7) |
| Published | 2026-4-13 |
| DOI | 10.1093/nar/gkag342 |
| PII | 8659121 |
| PMID | 42003550 |
| PMC | PMC13092967 |
| MeSH | Bacterial Proteins CRISPR-Associated Proteins* / genetics CRISPR-Associated Proteins* / metabolism CRISPR-Cas Systems* Deoxyribonucleases, Type II Site-Specific* / chemistry Deoxyribonucleases, Type II Site-Specific* / genetics Deoxyribonucleases, Type II Site-Specific* / metabolism Endodeoxyribonucleases* / genetics Endodeoxyribonucleases* / metabolism Gene Editing* / methods HEK293 Cells Humans Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism |
| IF | 11.502 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 5 |
| Altmetric score changes over past 6months | 2.7 |
| Resource | |
| Human and Animal Cells | K562(RCB1897) |