Rapid tagging of endogenous mouse genes by recombineering and ES cell complementation of tetraploid blastocysts

Nucleic Acids Research
Dewang ZhouTim M Townes

Abstract

The construction of knockin vectors designed to modify endogenous genes in embryonic stem (ES) cells and the generation of mice from these modified cells is time consuming. The timeline of an experiment from the conception of an idea to the availability of mature mice is at least 9 months. We describe a method in which this timeline is typically reduced to 3 months. Knockin vectors are rapidly constructed from bacterial artificial chromosome clones by recombineering followed by gap-repair (GR) rescue, and mice are rapidly derived by injecting genetically modified ES cells into tetraploid blastocysts. We also describe a tandem affinity purification (TAP)/floxed marker gene plasmid and a GR rescue plasmid that can be used to TAP tag any murine gene. The combination of recombineering and tetraploid blastocyst complementation provides a means for large-scale TAP tagging of mammalian genes.

References

Jan 27, 1995·The Journal of Biological Chemistry·D DonzeJ J Bieker
Sep 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·A NagyJ C Roder
May 17, 2000·Proceedings of the National Academy of Sciences of the United States of America·D YuD L Court
Nov 28, 2000·The Journal of Biological Chemistry·K PandyaT M Townes
May 2, 2001·Proceedings of the National Academy of Sciences of the United States of America·K EgganR Jaenisch
Oct 26, 2002·Science·Tong Ihn LeeRichard A Young

❮ Previous
Next ❯

Citations

Mar 23, 2011·Biotechnology Letters·Yifeng Li
Mar 1, 2012·Proceedings of the National Academy of Sciences of the United States of America·Yi-Shin LaiTim M Townes
Jan 24, 2007·Molecular and Cellular Biology·Yu-Chiau ShyuChe-Kun James Shen
Dec 10, 2015·Stem Cells and Development·Areta M CzerwinskaMaria Anna Ciemerych
Dec 22, 2004·The Journal of Physiology·Anne-Claude GingrasBrian Raught
Jan 13, 2016·Cellular and Molecular Life Sciences : CMLS·Yan ZhouYonglun Luo
Mar 19, 2016·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Giordanni C DantasHenrique Ferreira
Mar 11, 2008·Experimental Cell Research·Karen J QuadriniJames J Bieker
May 23, 2012·Proteomics·Wade H DunhamAnne-Claude Gingras
Sep 4, 2010·Biology of the Cell·Serge HardyLuc Paillard
Apr 26, 2016·Scientific Reports·Matthew J BroadheadSeth G N Grant
Jun 24, 2006·Methods in Enzymology·Ulrich Rass, Stephen C West

❮ Previous
Next ❯

Related Concepts

Related Feeds

Artificial Chromosomes

Artificial chromosomes are genetically engineered chromosomes derived from the DNA of a species. Discover the latest research on artificial chromosomes here.