Gene targeting using zinc finger nucleases

Abstract

The ability to achieve site-specific manipulation of the mammalian genome has widespread implications for basic and applied research. Gene targeting is a process in which a DNA molecule introduced into a cell replaces the corresponding chromosomal segment by homologous recombination, and thus presents a precise way to manipulate the genome. In the past, the application of gene targeting to mammalian cells has been limited by its low efficiency. Zinc finger nucleases (ZFNs) show promise in improving the efficiency of gene targeting by introducing DNA double-strand breaks in target genes, which then stimulate the cell's endogenous homologous recombination machinery. Recent results have shown that ZFNs can be used to create targeting frequencies of up to 20% in a human disease-causing gene. Future work will be needed to translate these in vitro findings to in vivo applications and to determine whether zinc finger nucleases create undesired genomic instability.

Knockdown of zinc finger protein X-linked inhibits prostate cancer cell proliferation and induces apoptosis by activating caspase-3 and caspase-9

“…6 There are a number of studies reporting diverse structures of zinc finger proteins corresponding to novel topologies. 6,7 Zinc finger protein X-linked (ZFX) is a member of the krueppel C 2 H 2 -type zinc-finger protein family and is on the X chromosome. This is structurally similar to a related gene on the Y chromosome, zinc finger protein Y-linked (ZFY).…”

Une nouvelle ère pour la génétique du rat

“…dans le gène ciblé. On sait depuis longtemps que, dans des systèmes cellulaires, l'induction ciblée d'une cassure double brin stimule localement les processus endogènes de réparation du génome et permet d'augmenter considéra-blement la fréquence de recombinaison homologue (voir par exemple [10][11][12][13]). La faisabilité de cette stratégie a récemment été démontrée chez la souris en co-injectant dans des zygotes des ARN codant pour des nucléases à doigts de zinc et une matrice de réparation présentant des séquences d'homologies de part et d'autre de la cassure.…”

Genome-Editing Technologies: Principles and Applications

“…ZFNs, which are fusions between a customdesigned Cys 2 -His 2 zinc-finger protein and the cleavage domain of the FokI restriction endonuclease (Kim et al 1996), were the first targeted nuclease to achieve widespread use (Porteus and Carroll 2005;Urnov et al 2010). ZFNs function as dimers, with each monomer recognizing a specific "half site" sequence-typically nine to 18 base pairs (bps) of DNA-via the zinc-finger DNA-binding domain (Fig.…”