Epigenetic Editing, Dept Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Hanzeplein 1, 9713 GZ Groningen, The Netherlands.
Chemical and Pharmaceutical Biology, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
Philos Trans R Soc Lond B Biol Sci. 2018 Jun 5;373(1748). doi: 10.1098/rstb.2017.0077.
The ability to target DNA specifically at any given position within the genome allows many intriguing possibilities and has inspired scientists for decades. Early gene-targeting efforts exploited chemicals or DNA oligonucleotides to interfere with the DNA at a given location in order to inactivate a gene or to correct mutations. We here describe an example towards correcting a genetic mutation underlying Pompe's disease using a nucleotide-fused nuclease (TFO-MunI). In addition to the promise of gene correction, scientists soon realized that genes could be inactivated or even re-activated without inducing potentially harmful DNA damage by targeting transcriptional modulators to a particular gene. However, it proved difficult to fuse protein effector domains to the first generation of programmable DNA-binding agents. The engineering of gene-targeting proteins (zinc finger proteins (ZFPs), transcription activator-like effectors (TALEs)) circumvented this problem. The disadvantage of protein-based gene targeting is that a fusion protein needs to be engineered for every locus. The recent introduction of CRISPR/Cas offers a flexible approach to target a (fusion) protein to the locus of interest using cheap designer RNA molecules. Many research groups now exploit this platform and the first human clinical trials have been initiated: CRISPR/Cas has kicked off a new era of gene targeting and is revolutionizing biomedical sciences.This article is part of a discussion meeting issue 'Frontiers in epigenetic chemical biology'.
靶向基因组中特定位置的 DNA 的能力为许多令人着迷的可能性提供了可能，并激发了科学家们几十年来的研究兴趣。早期的基因靶向研究利用化学物质或 DNA 寡核苷酸在特定位置干扰 DNA，以灭活基因或纠正突变。我们在这里描述了一个使用核苷酸融合核酸酶（TFO-MunI）纠正庞贝氏病遗传突变的例子。除了基因纠正的前景外，科学家们很快意识到，通过将转录调节剂靶向特定基因，可以在不引起潜在有害 DNA 损伤的情况下，使基因失活甚至重新激活。然而，将蛋白效应结构域融合到第一代可编程 DNA 结合剂中被证明是困难的。基因靶向蛋白（锌指蛋白（ZFPs）、转录激活因子样效应物（TALEs））的工程克服了这个问题。基于蛋白的基因靶向的缺点是，需要为每个基因座设计融合蛋白。最近引入的 CRISPR/Cas 提供了一种灵活的方法，使用廉价的设计 RNA 分子将（融合）蛋白靶向感兴趣的基因座。许多研究小组现在都在利用这个平台，并且已经启动了第一个人类临床试验：CRISPR/Cas 开启了基因靶向的新时代，正在彻底改变生物医学科学。本文是“表观遗传化学生物学前沿”讨论会议的一部分。
