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| Item Type: | Article |
|---|---|
| Title: | Protocol for efficient CRISPR/Cas9/AAV-mediated homologous recombination in mouse hematopoietic stem and progenitor cells |
| Creators Name: | Tran, N.T. and Trombke, J. and Rajewsky, K. and Chu, V.T. |
| Abstract: | Mutations that accumulate in self-renewing hematopoietic stem and progenitor cells (HSPCs) can cause severe blood disorders. To model such disorders in mice, we developed a CRISPR/Cas9/adeno-associated virus (AAV)-based system to knock in and repair genes by homologous recombination in mouse HSPCs. Here, we provide a step-by-step protocol to achieve high efficiency of gene knockin in mouse HSPCs, while maintaining engraftment capacity. This approach enables the functional study of hematopoietic disease mutations in vivo, without requiring germline mutagenesis. |
| Keywords: | CRISPR-Cas Systems, Gene Knock-In Techniques, Hematopoietic Stem Cells, Homologous Recombination, Stem Cells, Animals, Mice |
| Source: | STAR Protocols |
| ISSN: | 2666-1667 |
| Publisher: | Elsevier |
| Volume: | 1 |
| Number: | 1 |
| Page Range: | 100028 |
| Date: | 19 June 2020 |
| Official Publication: | https://doi.org/10.1016/j.xpro.2020.100028 |
| PubMed: | View item in PubMed |
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