Generation of a Xenopus laevis F1 albino J strain by genome editing and oocyte host-transfer

Dev Biol. 2017 Jun 15;426(2):188-193. doi: 10.1016/j.ydbio.2016.03.006. Epub 2016 Mar 15.

Abstract

Completion of the Xenopus laevis genome sequence from inbred J strain animals has facilitated the generation of germline mutant X. laevis using targeted genome editing. In the last few years, numerous reports have demonstrated that TALENs are able to induce mutations in F0 Xenopus embryos, but none has demonstrated germline transmission of such mutations in X. laevis. In this report we used the oocyte host-transfer method to generate mutations in both tyrosinase homeologs and found highly-penetrant germline mutations; in contrast, embryonic injections yielded few germline mutations. We also compared the distribution of mutations in several F0 somatic tissues and germ cells and found that the majority of mutations in each tissue were different. These results establish that X. laevis J strain animals are very useful for generating germline mutations and that the oocyte host-transfer method is an efficient technique for generating mutations in both homeologs.

Keywords: Genome editing; J strain; Oocyte host-transfer; TALENs; Tyrosinase; Xenopus laevis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Albinism / genetics*
  • Animals
  • Base Sequence
  • Crosses, Genetic
  • Embryo, Nonmammalian
  • Female
  • Gene Editing*
  • Germ-Line Mutation*
  • Male
  • Microinjections
  • Monophenol Monooxygenase / deficiency
  • Monophenol Monooxygenase / genetics*
  • Mosaicism
  • Oocytes / transplantation
  • Penetrance
  • RNA, Messenger / administration & dosage
  • RNA, Messenger / genetics
  • Sequence Alignment
  • Sequence Homology, Nucleic Acid
  • Transcription Activator-Like Effector Nucleases / genetics
  • Xenopus Proteins / deficiency
  • Xenopus Proteins / genetics*
  • Xenopus laevis / genetics*

Substances

  • RNA, Messenger
  • Xenopus Proteins
  • Monophenol Monooxygenase
  • Transcription Activator-Like Effector Nucleases