| First Author | Ueki K | Year | 2023 |
| Journal | Diabetes | Volume | 72 |
| Issue | 11 | Pages | 1609-1620 |
| PubMed ID | 37625131 | Mgi Jnum | J:343219 |
| Mgi Id | MGI:7564358 | Doi | 10.2337/db23-0445 |
| Citation | Ueki K, et al. (2023) Establishment of Pancreatic beta-Cell-Specific Gene Knockout System Based on CRISPR-Cas9 Technology With AAV8-Mediated gRNA Delivery. Diabetes 72(11):1609-1620 |
| abstractText | The Cre-loxP system provides valuable resources to analyze the importance of tissue-specific gene knockout (KO), including pancreatic beta-cells associated with the pathogenesis of diabetes. However, it is expensive and time consuming to generate transgenic mice harboring floxed genes of interest and cross them with cell-specific Cre expression mice. We establish a betaCas9 system with mice expressing Cas9 in pancreatic beta-cells and adeno-associated virus 8 (AAV8)-mediated guide RNA (gRNA) delivery based on CRISPR-Cas9 technology to overcome those shortcomings. Interbreeding CAG-loxP-STOP-loxP (LSL)-Cas9 with Ins1-Cre mice generates normal glucose-tolerant betaCas9 mice expressing Cas9 with fluorescent reporter EGFP specifically in beta-cells. We also show significant beta-cell-specific gene KO efficiency with AAV8-mediated delivery of gRNA for EGFP reporter by intraperitoneal injection in the mice. As a proof of concept, we administered AAV8 to betaCas9 mice for expressing gRNA for Pdx1, a culprit gene of maturity-onset diabetes of the young 4. As reported previously, we demonstrate that those mice show glucose intolerance with transdifferentiation of Pdx1 KO beta-cells into glucagon-expressing cells. We successfully generated a convenient beta-cell-specific gene KO system with betaCas9 mice and AAV8-mediated gRNA delivery. |
