- An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease.
An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease.
STAR protocols (2022-12-28)
Allen C T Teng, Marjan Tavassoli, Suja Shrestha, Ryan M Marks, Meghan J McFadden, Sonia L Evagelou, Kyle Lindsay, Ava Vandenbelt, Wenping Li, Evgueni Ivakine, Ronald Cohn, J Paul Santerre, Anthony O Gramolini
PMID36574341
ABSTRACT
Here, we describe a protocol for purifying functional clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) from Staphylococcus aureus within 24 h and over 90% purity. SaCas9 purification begins with immobilized metal affinity chromatography, followed by cation exchange chromatography, and ended with centrifugal concentrators. The simplicity, cost-effectiveness, and reproducibility of such protocols will enable general labs to produce a sizable amount of Cas9 proteins, further accelerating CRISPR research.