Affordable Access

deepdyve-link
Publisher Website

Generation of knockout mice using engineered nucleases

Authors
  • Sung, Young Hoon
  • Jin, Young
  • Kim, Seokjoong
  • Lee, Han-Woong1, 2, 3, 4, 5
  • 1 Department of Biochemistry
  • 2 College of Life Science and Biotechnology
  • 3 Laboratory Animal Research Center
  • 4 Yonsei University
  • 5 ToolGen, Inc.
Type
Published Article
Journal
Methods
Publication Date
Jan 01, 2014
Identifiers
DOI: 10.1016/j.ymeth.2014.02.009
Source
Elsevier
Keywords
License
Unknown

Abstract

The use of engineered nucleases in one-cell stage mouse embryos is emerging as an efficient alternative to conventional gene targeting in mouse embryonic stem (ES) cells. These nucleases are designed or reprogrammed to specifically induce double strand breaks (DSBs) at a desired genomic locus, and efficiently introduce mutations by both error-prone and error-free DNA repair mechanisms. Since these mutations frequently result in the loss or alteration of gene function by inserting, deleting, or substituting nucleotide sequences, engineered nucleases are enabling us to efficiently generate gene knockout and knockin mice. Three kinds of engineered endonucleases have been developed and successfully applied to the generation of mutant mice: zinc-finger nuclease (ZFNs), transcription activator-like effector nucleases (TALENs) and RNA-guided endonucleases (RGENs). Based on recent advances, here we provide experimentally validated, detailed guidelines for generating non-homologous end-joining (NHEJ)-mediated mutant mice by microinjecting TALENs and RGENs into the cytoplasm or the pronucleus of one-cell stage mouse embryos.

Report this publication

Statistics

Seen <100 times