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Improving the specificity of adenine base editor using high-fidelity Cas9
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ABSTRACT
Adenine base editor (ABE) mediates the conversion of A to G in genomic DNA. In human, approximately 47.8% of known pathogenic SNPs can be corrected by A to G conversion, indicating that ABE have tremendous potential in gene therapy. However, the off-target activity of ABE limits its clinical application. ABE off-target activity in DNA is depended on the bonding of
Streptococcus pyogenes
Cas9 (SpCas9) on off-target sites [1, 2]. Therefore, using high-fidelity Cas9 should be able to improve the specificity of ABE. Based on this, we replaced the wild-type SpCas9 in ABE7.10 with four high-fidelity Cas9s to improve its specificity. The analysis of target deep sequencing data demonstrate that the specificity of e-ABE is substantially improved compared to conventional ABE7.10 in four test sites. But the broad editing window of ABE hampers its application, ABE needs to be optimized to get variants with narrow editing window.
Title: Improving the specificity of adenine base editor using high-fidelity Cas9
Description:
ABSTRACT
Adenine base editor (ABE) mediates the conversion of A to G in genomic DNA.
In human, approximately 47.
8% of known pathogenic SNPs can be corrected by A to G conversion, indicating that ABE have tremendous potential in gene therapy.
However, the off-target activity of ABE limits its clinical application.
ABE off-target activity in DNA is depended on the bonding of
Streptococcus pyogenes
Cas9 (SpCas9) on off-target sites [1, 2].
Therefore, using high-fidelity Cas9 should be able to improve the specificity of ABE.
Based on this, we replaced the wild-type SpCas9 in ABE7.
10 with four high-fidelity Cas9s to improve its specificity.
The analysis of target deep sequencing data demonstrate that the specificity of e-ABE is substantially improved compared to conventional ABE7.
10 in four test sites.
But the broad editing window of ABE hampers its application, ABE needs to be optimized to get variants with narrow editing window.
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