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Texas Children's Hospital & Houston Methodist Hospital

1 ARTICLES PUBLISHED IN JoVE

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Genetics

Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9
Lorenzo Brunetti *1,2,3, Michael C. Gundry *1,2,4, Ayumi Kitano 4, Daisuke Nakada 1,2,4, Margaret A. Goodell 1,2,4,5
1Stem Cells & Regenerative Medicine Center, Baylor College of Medicine, 2Center for Cell and Gene Therapy, Baylor College of Medicine, 3Centro di Ricerca Emato-Oncologica (CREO), University of Perugia, 4Department of Molecular & Human Genetics, Baylor College of Medicine, 5Texas Children's Hospital & Houston Methodist Hospital

A protocol for fast CRISPR/Cas9-mediated gene disruption in mouse and human primary hematopoietic cells is described in this article. Cas9-sgRNA ribonucleoproteins are introduced via electroporation with sgRNAs generated through in vitro transcription and commercial Cas9. High editing efficiencies are achieved with limited time and financial cost.

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