Highly efficient and specific genome editing in silkworm using custom TALENs

PLoS One. 2012;7(9):e45035. doi: 10.1371/journal.pone.0045035. Epub 2012 Sep 18.

Abstract

Establishment of efficient genome editing tools is essential for fundamental research, genetic engineering, and gene therapy. Successful construction and application of transcription activator-like effector nucleases (TALENs) in several organisms herald an exciting new era for genome editing. We describe the production of two active TALENs and their successful application in the targeted mutagenesis of silkworm, Bombyx mori, whose genetic manipulation methods are parallel to those of Drosophila and other insects. We will also show that the simultaneous expression of two pairs of TALENs generates heritable large chromosomal deletion. Our results demonstrate that (i) TALENs can be used in silkworm and (ii) heritable large chromosomal deletions can be induced by two pairs of TALENs in whole organisms. The generation and the high frequency of TALENs-induced targeted mutagenesis in silkworm will promote the genetic modification of silkworm and other insect species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Bombyx / enzymology*
  • Bombyx / genetics*
  • Chromosome Deletion
  • Endonucleases / metabolism*
  • Female
  • Genetic Techniques*
  • Genetic Vectors / genetics
  • Genome, Insect / genetics*
  • Germ-Line Mutation / genetics
  • Molecular Sequence Data
  • Mosaicism
  • Mutagenesis / genetics
  • Phenotype
  • Trans-Activators / metabolism*

Substances

  • Trans-Activators
  • Endonucleases

Grant support

This work was supported by the Chinese Ministry of Science and Technology [2012CB114600, 2011AA100306] (http://www.most.gov.cn/); and the doctoral Innovation Fund of Southwest University [kb2010002, ky2009002] (http://www.swu.edu.cn/index.html). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.