| Autor: |
Jordi van Gestel, John S. Hawkins, Horia Todor, Carol A. Gross |
| Jazyk: |
angličtina |
| Rok vydání: |
2021 |
| Předmět: |
|
| Zdroj: |
STAR Protocols, Vol 2, Iss 2, Pp 100521- (2021) |
| Druh dokumentu: |
article |
| ISSN: |
2666-1667 |
| DOI: |
10.1016/j.xpro.2021.100521 |
| Popis: |
Summary: CRISPR interference is an increasingly popular method for perturbing gene expression. Guided by single-guide RNAs (sgRNAs), nuclease-deficient Cas9 proteins bind to specific DNA sequences and hinder transcription. Specificity is achieved through complementarity of the sgRNAs to the DNA. Changing complementarity by introducing single-nucleotide mismatches can be exploited to tune knockdown. Here, we present a computational pipeline to identify sgRNAs targeting specific genes in a bacterial genome, filter them, and titrate their activity by introducing mismatches.For complete details on the use and execution of this protocol, please refer to Hawkins et al. (2020). |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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