Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor Cells
Summary: 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 mous...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-06-01
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| Series: | STAR Protocols |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166720300150 |
| _version_ | 1811294826803494912 |
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| author | Ngoc Tung Tran Janine Trombke Klaus Rajewsky Van Trung Chu |
| author_facet | Ngoc Tung Tran Janine Trombke Klaus Rajewsky Van Trung Chu |
| author_sort | Ngoc Tung Tran |
| collection | DOAJ |
| description | Summary: 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.For complete details on the use and execution of this protocol, please refer to Tran et al. (2019). |
| first_indexed | 2024-04-13T05:23:16Z |
| format | Article |
| id | doaj.art-c5a008a78b1844ef9815fb8fe80112aa |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-13T05:23:16Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-c5a008a78b1844ef9815fb8fe80112aa2022-12-22T03:00:40ZengElsevierSTAR Protocols2666-16672020-06-0111100028Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor CellsNgoc Tung Tran0Janine Trombke1Klaus Rajewsky2Van Trung Chu3Immune Regulation and Cancer, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin 13125, GermanyImmune Regulation and Cancer, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin 13125, GermanyImmune Regulation and Cancer, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin 13125, GermanyImmune Regulation and Cancer, Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin 13125, Germany; Corresponding authorSummary: 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.For complete details on the use and execution of this protocol, please refer to Tran et al. (2019).http://www.sciencedirect.com/science/article/pii/S2666166720300150 |
| spellingShingle | Ngoc Tung Tran Janine Trombke Klaus Rajewsky Van Trung Chu Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor Cells STAR Protocols |
| title | Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor Cells |
| title_full | Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor Cells |
| title_fullStr | Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor Cells |
| title_full_unstemmed | Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor Cells |
| title_short | Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor Cells |
| title_sort | protocol for efficient crispr cas9 aav mediated homologous recombination in mouse hematopoietic stem and progenitor cells |
| url | http://www.sciencedirect.com/science/article/pii/S2666166720300150 |
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