| Autor: |
Adrian Pascal Nievergelt, Dennis Ray Diener, Aliona Bogdanova, Thomas Brown, Gaia Pigino |
| Jazyk: |
angličtina |
| Rok vydání: |
2024 |
| Předmět: |
|
| Zdroj: |
STAR Protocols, Vol 5, Iss 1, Pp 102774- (2024) |
| Druh dokumentu: |
article |
| ISSN: |
2666-1667 |
| DOI: |
10.1016/j.xpro.2023.102774 |
| Popis: |
Summary: CRISPR-Cas genome engineering in the unicellular green algal model Chlamydomonas reinhardtii has until recently suffered from low integration efficiencies despite traditional genetics being well established. Here, we present a protocol for efficient homology-directed knockin mutagenesis in all commonly used strains of Chlamydomonas. We describe steps for scarless integration of fusion tags and sequence modifications of almost all proteins without the need for a preceding mutant line. We further empower this genetic-editing approach by efficient crossing and highly robust screening protocols.For complete details on the use and execution of this protocol, please refer to Nievergelt et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
|
