| Autor: |
Franziska, Wimmer, Frank, Englert, Chase L, Beisel |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
Methods in molecular biology (Clifton, N.J.). 2433 |
| ISSN: |
1940-6029 |
| Popis: |
Type I CRISPR-Cas systems represent the most common and diverse type of these prokaryotic defense systems and are being harnessed for a growing set of applications. As these systems rely on multi-protein effector complexes, their characterization remains challenging. Here, we report a rapid and straightforward method to characterize these systems in a cell-free transcription-translation (TXTL) system. A ribonucleoprotein complex is produced and binds to its target next to a recognized PAM, thereby preventing the targeted sequence from being cleaved by a restriction enzyme. Selection for uncleaved targeted plasmids leads to an enrichment of recognized sequences within a PAM library. This assay will aid the exploration of CRISPR-Cas diversity and evolution and help contribute new systems for CRISPR technologies and applications. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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