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Protocol for efficient CRISPR/Cas9/AAV-mediated homologous recombination in mouse hematopoietic stem and progenitor cells

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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., Trombke, J., 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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