Functional profiling of single CRISPR/Cas9-edited human long-term hematopoietic stem cells

Elvin Wagenblast, Maria Azkanaz, Sabrina A. Smith, Lorien Shakib, Jessica L. McLeod, Gabriela Krivdova, Joana Araújo, Leonard D. Shultz, Olga I. Gan, John E. Dick () and Eric R. Lechman ()
Additional contact information
Elvin Wagenblast: University Health Network
Maria Azkanaz: University Health Network
Sabrina A. Smith: University Health Network
Lorien Shakib: University Health Network
Jessica L. McLeod: University Health Network
Gabriela Krivdova: University Health Network
Joana Araújo: University Health Network
Leonard D. Shultz: The Jackson Laboratory
Olga I. Gan: University Health Network
John E. Dick: University Health Network
Eric R. Lechman: University Health Network

Nature Communications, 2019, vol. 10, issue 1, 1-11

Abstract: Abstract In the human hematopoietic system, rare self-renewing multipotent long-term hematopoietic stem cells (LT-HSCs) are responsible for the lifelong production of mature blood cells and are the rational target for clinical regenerative therapies. However, the heterogeneity in the hematopoietic stem cell compartment and variable outcomes of CRISPR/Cas9 editing make functional interrogation of rare LT-HSCs challenging. Here, we report high efficiency LT-HSC editing at single-cell resolution using electroporation of modified synthetic gRNAs and Cas9 protein. Targeted short isoform expression of the GATA1 transcription factor elicit distinct differentiation and proliferation effects in single highly purified LT-HSC when analyzed with functional in vitro differentiation and long-term repopulation xenotransplantation assays. Our method represents a blueprint for systematic genetic analysis of complex tissue hierarchies at single-cell resolution.

Date: 2019
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-12726-0

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DOI: 10.1038/s41467-019-12726-0

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