Efficient production of a gene mutant cell line through integrating TALENs and high-throughput cell cloning

J Lab Autom. 2015 Feb;20(1):46-50. doi: 10.1177/2211068214562628. Epub 2014 Dec 18.

Abstract

Transcription activator-like effectors (TALEs) are becoming powerful DNA-targeting tools in a variety of mammalian cells and model organisms. However, generating a stable cell line with specific gene mutations in a simple and rapid manner remains a challenging task. Here, we report a new method to efficiently produce monoclonal cells using integrated TALE nuclease technology and a series of high-throughput cell cloning approaches. Following this method, we obtained three mTOR mutant 293T cell lines within 2 months, which included one homozygous mutant line.

Keywords: TALEN; gene mutant; high-throughput; stable cell line.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cloning, Organism / methods*
  • Cytological Techniques / methods*
  • Gene Targeting / methods*
  • Genetic Engineering / methods*
  • High-Throughput Screening Assays
  • Humans