| Autor: |
Thaddeus D Seher, Namkha Nguyen, Diana Ramos, Priyanka Bapat, Clarissa J Nobile, Suzanne S Sindi, Aaron D Hernday |
| Jazyk: |
angličtina |
| Rok vydání: |
2021 |
| Předmět: |
|
| Zdroj: |
G3: Genes, Genomes, Genetics, Vol 11, Iss 9 (2021) |
| Druh dokumentu: |
article |
| ISSN: |
2160-1836 |
| DOI: |
10.1093/g3journal/jkab216 |
| Popis: |
AbstractCRISPR/Cas-induced genome editing is a powerful tool for genetic engineering, however, targeting constraints limit which loci are editable with this method. Since the length of a DNA sequence impacts the likelihood it overlaps a unique target site, precision editing of small genomic features with CRISPR/Cas remains an obstacle. We introduce a two-step genome editing strategy that virtually eliminates CRISPR/Cas targeting constraints and facilitates precision genome editing of elements as short as a single base-pair at virtually any locus in any organism that supports CRISPR/Cas-induced genome editing. Our two-step approach first replaces the locus of interest with an “AddTag” sequence, which is subsequently replaced with any engineered sequence, and thus circumvents the need for direct overlap with a unique CRISPR/Cas target site. In this study, we demonstrate the feasibility of our approach by editing transcription factor binding sites within Candida albicans |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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