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1,5-isoquinolinediol Increases the Frequency of Gene Targeting by Homologous Recombination in Mouse Fibroblasts

1,5-isoquinolinediol Increases the Frequency of Gene Targeting by Homologous Recombination in Mouse Fibroblasts

Alexandre Semionov, Denis Cournoyer, Terry Y K Chow

Biochem Cell Biol. 2003 Feb;81(1):17-24.

PMID: 12683632

Abstract:

Gene targeting is a technique that allows the introduction of predefined alterations into chromosomal DNA. It involves a homologous recombination reaction between the targeted genomic sequence and an exogenous targeting vector. In theory, gene targeting constitutes the ideal method of gene therapy for single gene disorders. In practice, gene targeting remains extremely inefficient for at least two reasons: very low frequency of homologous recombination in mammalian cells and high proficiency of the mammalian cells to randomly integrate the targeting vector by illegitimate recombination. One known method to improve the efficiency of gene targeting is inhibition of poly(ADP-ribose)polymerase (PARP). It has been shown that PARP inhibitors, such as 3-methoxybenzamide, could lower illegitimate recombination, thus increasing the ratio of gene targeting to random integration. However, the above inhibitors were reported to decrease the absolute frequency of gene targeting. Here we show that treatment of mouse Ltk cells with 1,5-isoquinolinediol, a recent generation PARP inhibitor, leads to an increase up to 8-fold in the absolute frequency of gene targeting in the correction of the mutation at the stable integrated HSV tk gene.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP5154029-A	1,5-Isoquinolinediol		5154-02-9	Price

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