Genome Editing of Silkworms

Methods Mol Biol. 2023:2637:359-374. doi: 10.1007/978-1-0716-3016-7_27.

Abstract

Silkworm is a lepidopteran insect that has been used as a model for a wide variety of biological studies. The microinjection technique is available, and it is possible to cause transgenesis as well as target gene disruption via the genome editing technique. TALEN-mediated knockout is especially effective in this species. We also succeeded in the precise and efficient integration of a donor vector using the precise integration into target chromosome (PITCh) method. Here we describe protocols for ZFN (zinc finger nuclease)-, TALEN (transcription activator-like effector nuclease)-, and CRISPR/Cas9-mediated genome editing as well as the PITCh technique in the silkworm. We consider that all of these techniques can contribute to the further promotion of various biological studies in the silkworm and other insect species.

Keywords: CRISPR/Cas9; Knock-in; PITCh; Silkworm; TALEN; ZFN.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bombyx* / genetics
  • Bombyx* / metabolism
  • CRISPR-Cas Systems / genetics
  • Gene Editing* / methods
  • Transcription Activator-Like Effector Nucleases / genetics
  • Transcription Activator-Like Effector Nucleases / metabolism
  • Zinc Finger Nucleases / genetics
  • Zinc Finger Nucleases / metabolism

Substances

  • Transcription Activator-Like Effector Nucleases
  • Zinc Finger Nucleases