Abstract
The microbial CRISPR-Cas adaptive immune system can be harnessed to facilitate genome editing in eukaryotic cells (Cong L et al., Science 339, 819–823, 2013; Mali P et al., Science 339, 823–826, 2013). Here we describe a protocol for the use of the RNA-guided Cas9 nuclease from the Streptococcus pyogenes type II CRISPR system to achieve specific, scalable, and cost-efficient genome editing in mammalian cells.
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Acknowledgments
We thank Randall Platt for comments and members of the Zhang Lab for discussion, support, and advice. N.P. is supported by the National Science Foundation Graduate Research Fellowship, Primary Award #1122374. P.D.H. is a James Mills Pierce Fellow. F.Z. is supported by the NIH Transformative R01 Award (R01-NS073124); the NIH Director’s Pioneer Award (DP1-MH100706); the Keck, McKnight, Gates, Damon Runyon, Searle Scholars, Merkin, Klingenstein, and Simons Foundations; Bob Metcalfe; Mike Boylan; and Jane Pauley. Sequence and reagent information are available through http://www.genome-engineering.org.
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Pyzocha, N.K., Ran, F.A., Hsu, P.D., Zhang, F. (2014). RNA-Guided Genome Editing of Mammalian Cells. In: Storici, F. (eds) Gene Correction. Methods in Molecular Biology, vol 1114. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-761-7_17
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DOI: https://doi.org/10.1007/978-1-62703-761-7_17
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