RAG2

Biology

Gene

Genome editing

Recombination

Genome

Retrovirus

Retrotransposon

Computational biology

Coding region

Transposable element

Genetics

Nature Communications

Cytosine base editors (CBEs) enable programmable C-to-T conversion without DNA double-stranded breaks and homology-directed repair in a variety of organisms, which exhibit great potential for agricultural and biomedical applications. However, all reported cases only involved C-to-T substitution at a single targeted genomic site. Whether C-to-T substitution is effective in multiple sites/loci has not been verified in large animals. Here, by using pigs, an important animal for agriculture and biomedicine, as the subjective animal, we showed that CBEs could efficiently induce C-to-T conversions at multiple sites/loci with the combination of three genes, including DMD , TYR , and LMNA , or RAG1 , RAG2 , and IL2RG , simultaneously, at the embryonic and cellular levels. CBEs also could disrupt genes ( pol gene of porcine endogenous retrovirus) with dozens of copies by introducing multiple premature stop codons. With the CBEs, pigs carrying single gene or multiple gene point mutations were generated through embryo injection or nuclear transfer approach.

Efficient base editing for multiple genes and loci in pigs using base editors