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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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Efficient base editing for multiple genes and loci in pigs using base editors.

Jingke Xie1,2,3,4, Weikai Ge1,2,3,4, Nan Li1,2,3,4

  • 1CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou Medical University, Guangzhou, 510530, China.

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|June 30, 2019
PubMed
Summary
This summary is machine-generated.

Cytosine base editors (CBEs) efficiently create multiple C-to-T changes in pigs, advancing agricultural and biomedical applications. This study demonstrates CBEs

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Area of Science:

  • Genetics and Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Cytosine base editors (CBEs) offer precise C-to-T conversion without DNA breaks.
  • Previous applications were limited to single genomic sites.
  • Effectiveness in large animals for multiplex editing remained unverified.

Purpose of the Study:

  • To investigate the efficacy of CBEs for simultaneous C-to-T conversion at multiple genomic sites in pigs.
  • To assess the potential of CBEs for gene disruption in multi-copy genes.
  • To generate pigs with single or multiple point mutations using CBEs.

Main Methods:

  • Application of CBEs in pig embryos and cells.
  • Targeting multiple genes (DMD, TYR, LMNA, RAG1, RAG2, IL2RG) simultaneously.
  • Utilizing embryo injection and nuclear transfer for generating mutant pigs.
  • Disruption of porcine endogenous retrovirus (PERV) pol gene.

Main Results:

  • Efficient C-to-T conversions were achieved at multiple loci in pig cells and embryos.
  • Simultaneous editing of multiple genes was successfully demonstrated.
  • Gene disruption via premature stop codons was confirmed in multi-copy genes.
  • Pigs with targeted single and multiple point mutations were generated.

Conclusions:

  • CBEs are effective for multiplex genomic editing in large animals like pigs.
  • This technology holds significant promise for agricultural and biomedical applications.
  • CBEs provide a versatile tool for generating genetically modified pigs with precise mutations.