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Temporal control of Cre recombinase-mediated in vitro DNA recombination by Tet-on gene expression system.
Acta Biochim Biophys Sin (Shanghai). 2005 Feb; 37(2):133-8.AB

Abstract

Conditional gene expression and gene deletion are important experimental approaches for examining the functions of particular gene products in mouse models. These strategies exploiting Cre-mediated site-specific DNA recombination have been incorporated into transgenic and gene-targeting procedures to allow in vivo manipulation of DNA in embryonic stem cells (ES cells) or living animals. The Cre/lox P system has become widely used in conditional gene targeting, conditional gene repair and activation, inducible chromosome translocation, and chromosome engineering. In this project, we have employed the universal transgenic system and the liver-specific promoter system for tightly temporal and liver-specific control of Cre gene expression in mice that (1) integrates the advantages of the Tet-on gene expression system and Cre/lox P site-mediated gene activation, and (2) simplifies the scheme of animal crosses through a combination of two control elements in a single transgene. A liver-specific apoE promoter was inserted into the promoter cloning site upstream of the rtTA cassette of pCore construct to generate the transgene construct pApoErtTA-tetO-Cre, followed by demonstrating stringent regulation of doxycycline (Dox)-induced Cre-mediated recombination in the lox P-flanked transcription STOP cassette-modified BEL-7402 cells. That is to say, in the absence of Dox, the Cre gene is not expressed and will not induce site-specific recombination between two lox P sites, whereas on exposure to Dox, the Cre gene will be expressed and the recombination will occur. Together, these data indicate that the Tet-on gene expression system is able to successfully and stringently control Cre expression in vitro, which lays a solid foundation for efficient and spatio-temporal Cre gene activation in transgenic mice.

Authors+Show Affiliations

Center of Experimental Animals, Sun Yat-Sen (Zhongshan) University, Guangzhou 510080, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15685371

Citation

Guo, Zhong-Min, et al. "Temporal Control of Cre Recombinase-mediated in Vitro DNA Recombination By Tet-on Gene Expression System." Acta Biochimica Et Biophysica Sinica, vol. 37, no. 2, 2005, pp. 133-8.
Guo ZM, Xu K, Yue Y, et al. Temporal control of Cre recombinase-mediated in vitro DNA recombination by Tet-on gene expression system. Acta Biochim Biophys Sin (Shanghai). 2005;37(2):133-8.
Guo, Z. M., Xu, K., Yue, Y., Huang, B., Deng, X. Y., Zhong, N. Q., Hong, X., Chen, X. G., & Xiao, D. (2005). Temporal control of Cre recombinase-mediated in vitro DNA recombination by Tet-on gene expression system. Acta Biochimica Et Biophysica Sinica, 37(2), 133-8.
Guo ZM, et al. Temporal Control of Cre Recombinase-mediated in Vitro DNA Recombination By Tet-on Gene Expression System. Acta Biochim Biophys Sin (Shanghai). 2005;37(2):133-8. PubMed PMID: 15685371.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Temporal control of Cre recombinase-mediated in vitro DNA recombination by Tet-on gene expression system. AU - Guo,Zhong-Min, AU - Xu,Kang, AU - Yue,Ying, AU - Huang,Bing, AU - Deng,Xin-Yan, AU - Zhong,Nü-Qi, AU - Hong,Xun, AU - Chen,Xi-Gu, AU - Xiao,Dong, PY - 2005/2/3/pubmed PY - 2005/8/4/medline PY - 2005/2/3/entrez SP - 133 EP - 8 JF - Acta biochimica et biophysica Sinica JO - Acta Biochim Biophys Sin (Shanghai) VL - 37 IS - 2 N2 - Conditional gene expression and gene deletion are important experimental approaches for examining the functions of particular gene products in mouse models. These strategies exploiting Cre-mediated site-specific DNA recombination have been incorporated into transgenic and gene-targeting procedures to allow in vivo manipulation of DNA in embryonic stem cells (ES cells) or living animals. The Cre/lox P system has become widely used in conditional gene targeting, conditional gene repair and activation, inducible chromosome translocation, and chromosome engineering. In this project, we have employed the universal transgenic system and the liver-specific promoter system for tightly temporal and liver-specific control of Cre gene expression in mice that (1) integrates the advantages of the Tet-on gene expression system and Cre/lox P site-mediated gene activation, and (2) simplifies the scheme of animal crosses through a combination of two control elements in a single transgene. A liver-specific apoE promoter was inserted into the promoter cloning site upstream of the rtTA cassette of pCore construct to generate the transgene construct pApoErtTA-tetO-Cre, followed by demonstrating stringent regulation of doxycycline (Dox)-induced Cre-mediated recombination in the lox P-flanked transcription STOP cassette-modified BEL-7402 cells. That is to say, in the absence of Dox, the Cre gene is not expressed and will not induce site-specific recombination between two lox P sites, whereas on exposure to Dox, the Cre gene will be expressed and the recombination will occur. Together, these data indicate that the Tet-on gene expression system is able to successfully and stringently control Cre expression in vitro, which lays a solid foundation for efficient and spatio-temporal Cre gene activation in transgenic mice. SN - 1672-9145 UR - https://www.unboundmedicine.com/medline/citation/15685371/Temporal_control_of_Cre_recombinase_mediated_in_vitro_DNA_recombination_by_Tet_on_gene_expression_system_ L2 - http://www.informatics.jax.org/reference/15685371 DB - PRIME DP - Unbound Medicine ER -