Highly efficient multiplex targeted mutagenesis and genomic structure variation in Bombyx mori cells using CRISPR/Cas9

Insect Biochem Mol Biol. 2014 Jun;49:35-42. doi: 10.1016/j.ibmb.2014.03.010. Epub 2014 Mar 31.

Abstract

Bombyx mori is an economically important insect and a model organism for studying lepidopteran and arthropod biology. Using a highly efficient CRISPR/Cas9 system, we showed that this system could mediate highly efficient targeted genome editing of a single gene locus, large chromosomal deletion or inversion, and also multiplex genome editing of 6 genes simultaneously in BmNs cell line derived from B. mori. The simplicity and high efficiency of our system provide unprecedented possibilities for researchers to implement precise and sophisticated manipulation of a chosen B. mori gene in BmNs cells easily in a limited time course, and perhaps new opportunities for functional genomics of B. mori and other lepidopteran insects.

Keywords: Bombyx mori; CRISPR/Cas9; Chromosomal deletion; Chromosomal inversion; Multiplex genome editing; Mutagenesis.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bombyx / genetics*
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Gene Targeting / methods*
  • Genetic Variation
  • Genomics
  • Insect Proteins / genetics
  • Mutagenesis

Substances

  • Insect Proteins