Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 22, Issue 8, Pages -Publisher
MDPI
DOI: 10.3390/ijms22083834
Keywords
CRISPR; Cas; homology directed repair; DNA repair; synthesis-dependent strand annealing; gene repair
Funding
- National Institute of General Medical Sciences [P20GM109021, P20GM103446]
- Nemours/B + Foundation [DDD604515]
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The mechanism of ssODN-directed gene editing has been a topic of discussion within the field of CRISPR gene editing. The ExACT pathway, based on oligo-driven DNA repair, provides a comprehensive understanding of the different ways DNA repair can occur in the presence of a repair oligonucleotide after CRISPR cleavage, by comparing and challenging other similar DNA repair pathways.
The mechanism of action of ssODN-directed gene editing has been a topic of discussion within the field of CRISPR gene editing since its inception. Multiple comparable, but distinct, pathways have been discovered for DNA repair both with and without a repair template oligonucleotide. We have previously described the ExACT pathway for oligo-driven DNA repair, which consisted of a two-step DNA synthesis-driven repair catalyzed by the simultaneous binding of the repair oligonucleotide (ssODN) upstream and downstream of the double-strand break. In order to better elucidate the mechanism of ExACT-based repair, we have challenged the assumptions of the pathway with those outlines in other similar non-ssODN-based DNA repair mechanisms. This more comprehensive iteration of the ExACT pathway better described the many different ways where DNA repair can occur in the presence of a repair oligonucleotide after CRISPR cleavage, as well as how these previously distinct pathways can overlap and lead to even more unique repair outcomes.
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