Generation of sheep with defined FecBB and TBXT mutations and porcine blastocysts with KCNJ5G151R/+ mutation using prime editing

Shiwei Zhou, Laura Johanna Lenk, Yawei Gao, Yuhui Wang, Xiaoe Zhao, Menghao Pan, Shuhong Huang, Kexin Sun, Peter Kalds, Qi Luo, Simon Lillico, Tad Sonstegard, Ute I Scholl, Baohua Ma, Bjoern Petersen, Yulin Chen, Xiaolong Wang

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

BACKGROUND: Rewriting the genomes of living organisms has been a long-standing aim in the biological sciences. The revelation of the CRISPR/Cas9 technology has revolutionized the entire biological field. Since its emergence, this technology has been widely applied to induce gene knockouts, insertions, deletions, and base substitutions. However, the classical version of this system was imperfect for inducing or correcting desired mutations. A subsequent development generated more advanced classes, including cytosine and adenine base editors, which can be used to achieve single nucleotide substitutions. Nevertheless, these advanced systems still suffer from several limitations, such as the inability to edit loci without a suitable PAM sequence and to induce base transversions. On the other hand, the recently emerged prime editors (PEs) can achieve all possible single nucleotide substitutions as well as targeted insertions and deletions, which show promising potential to alter and correct the genomes of various organisms. Of note, the application of PE to edit livestock genomes has not been reported yet.

RESULTS: In this study, using PE, we successfully generated sheep with two agriculturally significant mutations, including the fecundity-related FecBB p.Q249R and the tail length-related TBXT p.G112W. Additionally, we applied PE to generate porcine blastocysts with a biomedically relevant point mutation (KCNJ5 p.G151R) as a porcine model of human primary aldosteronism.

CONCLUSIONS: Our study demonstrates the potential of the PE system to edit the genomes of large animals for the induction of economically desired mutations and for modeling human diseases. Although prime-edited sheep and porcine blastocysts could be generated, the editing frequencies are still unsatisfactory, highlighting the need for optimizations in the PE system for efficient generation of large animals with customized traits.

Original languageEnglish
Article number313
Pages (from-to)1-9
Number of pages9
JournalBMC Genomics
Volume24
Issue number1
Early online date12 Jun 2023
DOIs
Publication statusE-pub ahead of print - 12 Jun 2023

Keywords / Materials (for Non-textual outputs)

  • Humans
  • Animals
  • Swine
  • Sheep
  • Mutation
  • Point Mutation
  • Blastocyst
  • Livestock
  • Nucleotides
  • G Protein-Coupled Inwardly-Rectifying Potassium Channels

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