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Randomized PAM depletion of phylogenetically-diverse Cas12a nucleases

ABSTRACT: We report the PAMs of phylogenetically-diverse Cas12a nucleases using a cell-free TXTL-based PAM screen. By adding a 5N randomized PAM library and Cas12a-gRNA in vitro, recognized PAM sequences were cleaved, while non-recognized PAMs remained. By amplifying the non-cleaved DNA, we used next-generation sequencing to analyze the depletion of functional PAMs of these Cas12a nucleases.

ORGANISM(S): synthetic construct

PROVIDER: GSE130377 | GEO | 2020/03/02

REPOSITORIES: GEO

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