Efficient production of Cre-mediated site-directed recombinants through the utilization of the puromycin resistance gene, pac: a transient gene-integration marker for ES cells - PubMed (original) (raw)

Efficient production of Cre-mediated site-directed recombinants through the utilization of the puromycin resistance gene, pac: a transient gene-integration marker for ES cells

M Taniguchi et al. Nucleic Acids Res. 1998.

Abstract

Gene targeting in embryonic stem (ES) cells is a powerful tool for generating mice carrying specifically designed mutations in the germline. Puromycin can completely kill ES cells within 24 to 48 h whereas G418 and hygromycin cannot. We have, therefore, proposed that the puromycin N-acetyltransferase ( pac ) gene, may be utilized as a transient gene-integration marker. Using a circular expression vector of cre and pac genes, Cre-mediated mutant cells were effectively enriched by pulse treatment of puromycin without stable integration of their genes. We have thus demonstrated the first application of pac as a transient gene-integration marker for ES cells.

PubMed Disclaimer

Similar articles

• Puromycin resistance gene as an effective selection marker for ciliate Tetrahymena.
  Iwamoto M, Mori C, Hiraoka Y, Haraguchi T. Iwamoto M, et al. Gene. 2014 Jan 25;534(2):249-55. doi: 10.1016/j.gene.2013.10.049. Epub 2013 Nov 1. Gene. 2014. PMID: 24185080
• Stable production of mutant mice from double gene converted ES cells with puromycin and neomycin.
  Watanabe S, Kai N, Yasuda M, Kohmura N, Sanbo M, Mishina M, Yagi T. Watanabe S, et al. Biochem Biophys Res Commun. 1995 Aug 4;213(1):130-7. doi: 10.1006/bbrc.1995.2107. Biochem Biophys Res Commun. 1995. PMID: 7639726
• Conditional mutagenesis in mice: the Cre/loxP recombination system.
  Plück A. Plück A. Int J Exp Pathol. 1996 Dec;77(6):269-78. Int J Exp Pathol. 1996. PMID: 9155661 Free PMC article. Review. No abstract available.
• [Modification of gene targeting method for functional analysis of the target gene in vivo].
  Shibata H, Kanamaru R, Noda T. Shibata H, et al. Gan To Kagaku Ryoho. 1997 Feb;24(4):460-5. Gan To Kagaku Ryoho. 1997. PMID: 9063484 Review. Japanese.

Cited by

• Increasing Knockin Efficiency in Mouse Zygotes by Transient Hypothermia.
  Bouchareb A, Biggs D, Alghadban S, Preece C, Davies B. Bouchareb A, et al. CRISPR J. 2024 Apr;7(2):111-119. doi: 10.1089/crispr.2023.0077. CRISPR J. 2024. PMID: 38635329 Free PMC article.
• Single-gene imaging links genome topology, promoter-enhancer communication and transcription control.
  Li J, Hsu A, Hua Y, Wang G, Cheng L, Ochiai H, Yamamoto T, Pertsinidis A. Li J, et al. Nat Struct Mol Biol. 2020 Nov;27(11):1032-1040. doi: 10.1038/s41594-020-0493-6. Epub 2020 Sep 21. Nat Struct Mol Biol. 2020. PMID: 32958948 Free PMC article.
• Conditional kisspeptin neuron-specific Kiss1 knockout with newly generated Kiss1-floxed and Kiss1-Cre mice replicates a hypogonadal phenotype of global Kiss1 knockout mice.
  Ikegami K, Goto T, Nakamura S, Watanabe Y, Sugimoto A, Majarune S, Horihata K, Nagae M, Tomikawa J, Imamura T, Sanbo M, Hirabayashi M, Inoue N, Maeda KI, Tsukamura H, Uenoyama Y. Ikegami K, et al. J Reprod Dev. 2020 Aug 20;66(4):359-367. doi: 10.1262/jrd.2020-026. Epub 2020 Apr 20. J Reprod Dev. 2020. PMID: 32307336 Free PMC article.
• The Small GTPase Rac1 Increases Cell Surface Stiffness and Enhances 3D Migration Into Extracellular Matrices.
  Kunschmann T, Puder S, Fischer T, Steffen A, Rottner K, Mierke CT. Kunschmann T, et al. Sci Rep. 2019 May 22;9(1):7675. doi: 10.1038/s41598-019-43975-0. Sci Rep. 2019. PMID: 31118438 Free PMC article.
• Transient AID expression for in situ mutagenesis with improved cellular fitness.
  Al-Qaisi TS, Su YC, Roffler SR. Al-Qaisi TS, et al. Sci Rep. 2018 Jun 20;8(1):9413. doi: 10.1038/s41598-018-27717-2. Sci Rep. 2018. PMID: 29925928 Free PMC article.

References

1. 1. Biochem Biophys Res Commun. 1995 Aug 4;213(1):130-7 - PubMed
2. 1. Cell. 1993 Jun 18;73(6):1155-64 - PubMed
3. 1. Anal Biochem. 1993 Oct;214(1):70-6 - PubMed
4. 1. Nat Genet. 1995 Apr;9(4):376-85 - PubMed
5. 1. Nucleic Acids Res. 1997 Aug 15;25(16):3326-31 - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Europe PubMed Central
  • PubMed Central
  • Silverchair Information Systems

• Other Literature Sources

  • The Lens - Patent Citations Database

• Medical

  • MedlinePlus Health Information