Article
Continuous evolution of SpCas9 variants compatible with non-G PAMs.
Nature biotechnology - 1 Apr 2020
Miller Shannon M, Wang Tina, Randolph Peyton B, Arbab Mandana, Shen Max W, Huang Tony P, Matuszek Zaneta, Newby Gregory A, Rees Holly A, Liu David R
Abstract excerpt
The targeting scope of Streptococcus pyogenes Cas9 (SpCas9) and its engineered variants is largely restricted to protospacer-adjacent motif (PAM) sequences containing G bases. Here we report the evolution of three new SpCas9 variants that collectively recognize NRNH PAMs (where R is A or G and H is A, C or T) using phage-assisted non-continuous evolution, three new phage-assisted continuous evolution strategies...
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