Abstract
Cytidine base editors (CBEs) and adenine base editors (ABEs), composed of a cytidine deaminase or an evolved adenine deaminase fused to Cas9 nickase, enable the conversion of C·G to T·A or A·T to G·C base pair in organisms, respectively. Here, we show that BE3 and ABE7.10 systems can achieve a targeted mutation efficiency of 53–88% and 44–100%, respectively, in both blastocysts and Founder (F0) rabbits. Meanwhile, this strategy can be used to precisely mimic human pathologies by efficiently inducing nonsense or missense mutations as well as RNA mis-splicing in rabbit. In addition, the reduced frequencies of indels with higher product purity are also determined in rabbit blastocysts by BE4-Gam, which is an updated version of the BE3 system. Collectively, this work provides a simple and efficient method for targeted point mutations and generation of disease models in rabbit.
Highlights
Cytidine base editors (CBEs) and adenine base editors (ABEs), composed of a cytidine deaminase or an evolved adenine deaminase fused to Cas[9] nickase, enable the conversion of C·G to T·A or A·T to G·C base pair in organisms, respectively
We firstly demonstrate that base editors provide a simple and highly efficient method for inducing single-nucleotide substitution in rabbit
No off-target mutations were detectable at potential off-target sites (POTs) in these loci of mutant blastocysts (Supplementary Fig. 10). These results suggested that the ABE7.10 system is highly efficient and precise in the conversion of the A·T to G·C base pair in rabbit blastocysts
Summary
Cytidine base editors (CBEs) and adenine base editors (ABEs), composed of a cytidine deaminase or an evolved adenine deaminase fused to Cas[9] nickase, enable the conversion of C·G to T·A or A·T to G·C base pair in organisms, respectively. We show that BE3 and ABE7.10 systems can achieve a targeted mutation efficiency of 53–88% and 44–100%, respectively, in both blastocysts and Founder (F0) rabbits. This work provides a simple and efficient method for targeted point mutations and generation of disease models in rabbit. The results provide a prospective application for the generation of rabbit models which could precisely mimic human genetic diseases. This is the first report, to our knowledge, demonstrating the reduction of frequencies of indels and improvement of product purity of the new BE version (BE4-Gam) in blastocysts, compared with BE3. We firstly demonstrate that base editors (both CBEs and ABEs) provide a simple and highly efficient method for inducing single-nucleotide substitution in rabbit
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