論文

国際誌
2021年2月23日

Lipofection-Mediated Introduction of CRISPR/Cas9 System into Porcine Oocytes and Embryos.

Animals : an open access journal from MDPI
  • Maki Hirata
  • ,
  • Manita Wittayarat
  • ,
  • Zhao Namula
  • ,
  • Quynh Anh Le
  • ,
  • Qingyi Lin
  • ,
  • Koki Takebayashi
  • ,
  • Chommanart Thongkittidilok
  • ,
  • Fuminori Tanihara
  • ,
  • Takeshige Otoi

11
2
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.3390/ani11020578

Liposome-mediated gene transfer has become an alternative method for establishing a gene targeting framework, and the production of mutant animals may be feasible even in laboratories without specialized equipment. However, how this system functions in mammalian oocytes and embryos remains unclear. The present study was conducted to clarify whether blastocyst genome editing can be performed by treatment with lipofection reagent, guide RNA, and Cas9 for 5 h without using electroporation or microinjection. A mosaic mutation was observed in blastocysts derived from zona pellucida (ZP)-free oocytes following lipofection treatment, regardless of the target genes. When lipofection treatment was performed after in vitro fertilization (IVF), no significant differences in the mutation rates or mutation efficiency were found between blastocysts derived from embryos treated at 24 and 29 h from the start of IVF. Only blastocysts from embryos exposed to lipofection treatment at 29 h after IVF contained biallelic mutant. Furthermore, there were no significant differences in the mutation rates or mutation efficiency between blastocysts derived from embryos at the 2- and 4-cell stages. This suggests that lipofection-mediated gene editing can be performed in ZP-free oocytes and ZP-free embryos; however, other factors affecting the system efficiency should be further investigated.

リンク情報
DOI
https://doi.org/10.3390/ani11020578
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/33672168
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7926877
ID情報
  • DOI : 10.3390/ani11020578
  • PubMed ID : 33672168
  • PubMed Central 記事ID : PMC7926877

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