Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels.

Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels. Research Abstract Details 

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Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels. Abstract Text:

Kai Liu,Susan Hipkens,Tao Yang,Robert Abraham,Wei Zhang,Nagesh Chopra,Bjorn Knollmann,Mark A Magnuson,Dan M Roden,

SCN5A encodes the predominant voltage-gated sodium channel isoform in human heart and nearly 100 variants have now been described and studied in vitro. However, development of animal models to analyze function of such large numbers of human gene variants represents a continuing challenge in translational medicine. Here, we describe the implementation of a two stage procedure, recombinase-mediated cassette exchange (RMCE), to efficiently and rapidly generate mice in which a full-length human cDNA replaces expression of the murine ortholog. In the first step of RMCE, conventional homologous recombination in mouse ES cells was used to replace scn5a exon 2 (that contains the translation start site) with a cassette acceptor that includes the thymidine kinase gene, flanked by loxP/inverted loxP sites. In the second step, the cassette acceptor site was replaced by the full-length wild-type human SCN5A cDNA by Cre/loxP-mediated recombination. The exchange event occurred in 7/29 (24%) colonies, and the time from electroporation to first homozygotes was only 8 months. PCR-restriction fragment length polymorphism (RFLP) showed that the murine isoform was replaced by the human one, and functional studies indicated that mice with human cardiac sodium channels have wild-type sodium current density, action potential durations, heart rates, and QRS durations. These data demonstrate that RMCE can be used to generate mice in which a targeted allele can be rapidly and efficiently replaced by variants of choice, and thereby can serve as an enabling approach for the functional characterization of ion channel and other DNA variants.

Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels. Publishing Authors By Initials

K Liu,S Hipkens,T Yang,R Abraham,W Zhang,N Chopra,B Knollmann,MA Magnuson,DM Roden,

For similar proteins: carrier proteins: membrane transport proteins: ion channels: sodium channels research abstracts see: proteins: carrier proteins: membrane transport proteins: ion channels: sodium channels research

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Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Genesis (New York, N.Y. : 2000)

VOLUME: 44

Page Numbers: 556-64

Journal Abbreviation: Genesis

ISSN: 1526-954X

DAY: 3

MONTH: Nov

YEAR: 2006

Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 100931242

Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels. Keywords Mesh Terms:

KEYWORDS: Sodium Channels

MESH TERMS: metabolism

Chemical & Substance for Abstract: Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels. Information

Substance Name: Integrases

Registry Number: EC 2.7.7.-

Grant and Affiliation Information for Recombinase-mediated cassette exchange to rapidly and efficiently generate mice with human cardiac sodium channels.

AFFILIATION: Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-0575, USA.

Country: United States

AGENCY: United States NHLBI

GRANT: HL46681

ACRONYM: HL

MEDLINETA: Genesis

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