Guide Swap enables genome-scale pooled CRISPR–Cas9 screening in human primary cells
2018
CRISPR–
Cas9screening allows genome-wide interrogation of gene function. Currently, to achieve the high and uniform
Cas9expression desirable for screening, one needs to engineer stable and clonal
Cas9-expressing cells—an approach that is not applicable in human
primary cells. Guide Swap permits genome-scale pooled
CRISPR–
Cas9screening in human
primary cellsby exploiting the unexpected finding that editing by lentivirally delivered, targeted
guide RNAs(gRNAs) occurs efficiently when
Cas9is introduced in complex with nontargeting gRNA. We validated Guide Swap in depletion and enrichment screens in CD4+ T cells. Next, we implemented Guide Swap in a model of ex vivo hematopoiesis, and identified known and previously unknown regulators of CD34+ hematopoietic stem and progenitor cell (HSPC) expansion. We anticipate that this platform will be broadly applicable to other challenging cell types, and thus will enable discovery in previously inaccessible but biologically relevant human
primary cellsystems.
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