A detailed protocol for expression, purification, and activity determination of recombinant SaCas9.
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ABSTRACT:
SUBMITTER: Flottmann F
PROVIDER: S-EPMC8991255 | biostudies-literature | 2022 Jun
REPOSITORIES: biostudies-literature
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A detailed protocol for expression, purification, and activity determination of recombinant SaCas9.
Flottmann Franziska F Pohl Greta Marie GM Gummert Jan J Milting Hendrik H Brodehl Andreas A
STAR protocols 20220405 2
Recombinant SaCas9 is useful for a broad range of applications in the context of genome editing, especially when the specific protospacer adjacent motifs of other Cas9 derivatives are missing. Here, we describe a detailed protocol for the expression and purification of recombinant SaCas9. We detail the main steps for immobilized metal affinity chromatography and size exclusion chromatography. In addition, we present an assay for activity determination of SaCas9. Active SaCas9 can be purified in ...[more]
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