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An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease
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 cati...
| Autores principales: | , , , , , , , , , , , , |
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| Formato: | Online Artículo Texto |
| Lenguaje: | English |
| Publicado: |
Elsevier
2022
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| Materias: | |
| Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9813775/ https://www.ncbi.nlm.nih.gov/pubmed/36574341 http://dx.doi.org/10.1016/j.xpro.2022.101933 |
| _version_ | 1784863993606701056 |
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| author | Teng, Allen C.T. Tavassoli, Marjan Shrestha, Suja Marks, Ryan M. McFadden, Meghan J. Evagelou, Sonia L. Lindsay, Kyle Vandenbelt, Ava Li, Wenping Ivakine, Evgueni Cohn, Ronald Santerre, J. Paul Gramolini, Anthony O. |
| author_facet | Teng, Allen C.T. Tavassoli, Marjan Shrestha, Suja Marks, Ryan M. McFadden, Meghan J. Evagelou, Sonia L. Lindsay, Kyle Vandenbelt, Ava Li, Wenping Ivakine, Evgueni Cohn, Ronald Santerre, J. Paul Gramolini, Anthony O. |
| author_sort | Teng, Allen C.T. |
| collection | PubMed |
| description | 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. |
| format | Online Article Text |
| id | pubmed-9813775 |
| institution | National Center for Biotechnology Information |
| language | English |
| publishDate | 2022 |
| publisher | Elsevier |
| record_format | MEDLINE/PubMed |
| spelling | pubmed-98137752023-01-06 An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease Teng, Allen C.T. Tavassoli, Marjan Shrestha, Suja Marks, Ryan M. McFadden, Meghan J. Evagelou, Sonia L. Lindsay, Kyle Vandenbelt, Ava Li, Wenping Ivakine, Evgueni Cohn, Ronald Santerre, J. Paul Gramolini, Anthony O. STAR Protoc Protocol 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. Elsevier 2022-12-26 /pmc/articles/PMC9813775/ /pubmed/36574341 http://dx.doi.org/10.1016/j.xpro.2022.101933 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
| spellingShingle | Protocol Teng, Allen C.T. Tavassoli, Marjan Shrestha, Suja Marks, Ryan M. McFadden, Meghan J. Evagelou, Sonia L. Lindsay, Kyle Vandenbelt, Ava Li, Wenping Ivakine, Evgueni Cohn, Ronald Santerre, J. Paul Gramolini, Anthony O. An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease |
| title | An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease |
| title_full | An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease |
| title_fullStr | An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease |
| title_full_unstemmed | An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease |
| title_short | An efficient and cost-effective purification protocol for Staphylococcus aureus Cas9 nuclease |
| title_sort | efficient and cost-effective purification protocol for staphylococcus aureus cas9 nuclease |
| topic | Protocol |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9813775/ https://www.ncbi.nlm.nih.gov/pubmed/36574341 http://dx.doi.org/10.1016/j.xpro.2022.101933 |
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