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17 April 2018
Yuichiro Miyaoka, PhD, (Regenerative Medicine Project) published a paper on “New way to enhance precise genome editing via HDR induced by CRISPR/Cas9.” in Nucleic Acids Research.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 with improved proof-reading enhances homology-directed repair

Background and Aim

CRISPR/Cas9 allows for genetic manipulations in basically all species, which has a great potential in therapeutic and agricultural applications. CRISPR/Cas9 specifically cuts target genomic DNA sequences to activate the two DNA repair pathways, NHEJ and HDR. NHEJ induces DNA insertions and deletions, whereas HDR precisely repairs the genome via recombination between the genomic DNA and template DNA. However, CRISPR/Cas9 always generates NHEJ along with HDR. Therefore, we investigated how the balance between HDR and NHEJ is determined by CRISPR/Cas9.

<Title of the paper>
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 with improved proof-reading enhances homology-directed repair
<Journal>
Nucleic Acids Research. Volume 46, Issue 9, 18 May 2018, Pages 4677–4688,
https://academic.oup.com/nar/advance-article/doi/10.1093/nar/gky264/4972875
Published: 17 April 2018

Methods and Results

We utilized our original methods to detect single nucleotide substitutions and knock-ins of a multi-kb gene fragment mediated by HDR, and found that Cas9 variants and modified gRNAs with altered interactions with the genomic DNA exhibited higher HDR/NHEJ ratios. Our results indicate that alterations in the interactions between CRISPR/Cas9 and the genomic DNA can lead to more precise and efficient genome editing mediated by HDR.


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