An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease
oleh: 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
| Format: | Article |
|---|---|
| Diterbitkan: | Elsevier 2023-03-01 |
Deskripsi
Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.