Gene Knockout in Hematopoietic Stem and Progenitor Cells Followed by Granulocytic Differentiation.

Perihan Mir, Malte Ritter, Karl Welte, Julia Skokowa, Maksim Klimiankou

Methods in molecular biology (Clifton, N.J.) 2020

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In this chapter, we present an optimized CRISPR/Cas9 RNP nucleofection approach for gene knockout (KO) in hematopoietic stem and progenitor cells (HSPCs). With experimentally proved active locus-specific sgRNAs, we routinely reach over 80% gene KO in HSPCs, thus avoiding the need for cell sorting or enrichment of targeted cell population. Additionally, we provide a protocol for in vitro granulocytic differentiation of HSPCs after gene KO and detailed description of granulocyte function tests which can be applied to study the effects of a particular gene KO.

Citation

Perihan Mir, Malte Ritter, Karl Welte, Julia Skokowa, Maksim Klimiankou. Gene Knockout in Hematopoietic Stem and Progenitor Cells Followed by Granulocytic Differentiation. Methods in molecular biology (Clifton, N.J.). 2020;2115:455-469