Institute of Nanotechnology, Karlsruhe Institute of Technology, 76021 Karlsruhe, Germany.
FEBS Lett. 2013 Oct 1;587(19):3231-5. doi: 10.1016/j.febslet.2013.08.015. Epub 2013 Aug 27.
Zinc finger nucleases are a promising tool to edit DNA in many biological applications, in particular for gene knockout. Despite many efforts the number of genes that can be effectively targeted with ZFNs remains severely limited, as available constructs cannot address arbitrary gene sequences. Here, we develop a novel concept to significantly enhance the number of DNA sequences that can be targeted by ZFN. Using an efficient computational model, we provide an extensive library of possible linker molecules between individual zinc finger motifs in the construct that can skip up to 10 base pairs between adjacent zinc finger recognition sites in the DNA sequence, which increases the number of genes that can be efficiently targeted by more than an order of magnitude.
锌指核酸酶是一种很有前途的工具，可用于许多生物学应用中的 DNA 编辑，特别是用于基因敲除。尽管已经做出了很多努力，但可用的构建体仍然无法解决任意基因序列，因此能够有效靶向的基因数量仍然严重受限。在这里，我们开发了一种新的概念，可以显著提高 ZFN 可以靶向的 DNA 序列数量。使用有效的计算模型，我们提供了一个可能的链接器分子的广泛库，这些分子可以在构建体中的单个锌指基序之间跳过多达 10 个碱基对，从而增加了可以有效靶向的基因数量，提高了一个数量级以上。
