Department of Molecular, Cell and Developmental Biology, University of California at Los Angeles, Los Angeles, California, USA.
Howard Hughes Medical Institute (HHMI), UCLA, Los Angeles, California, USA.
Plant Physiol. 2022 Mar 28;188(4):1811-1824. doi: 10.1093/plphys/kiac033.
Tools for sequence-specific DNA binding have opened the door to new approaches in investigating fundamental questions in biology and crop development. While there are several platforms to choose from, many of the recent advances in sequence-specific targeting tools are focused on developing Clustered Regularly Interspaced Short Palindromic Repeats- CRISPR Associated (CRISPR-Cas)-based systems. Using a catalytically inactive Cas protein (dCas), this system can act as a vector for different modular catalytic domains (effector domains) to control a gene's expression or alter epigenetic marks such as DNA methylation. Recent trends in developing CRISPR-dCas systems include creating versions that can target multiple copies of effector domains to a single site, targeting epigenetic changes that, in some cases, can be inherited to the next generation in the absence of the targeting construct, and combining effector domains and targeting strategies to create synergies that increase the functionality or efficiency of the system. This review summarizes and compares DNA targeting technologies, the effector domains used to target transcriptional control and epi-mutagenesis, and the different CRISPR-dCas systems used in plants.
序列特异性 DNA 结合工具为研究生物学和作物发展中的基本问题开辟了新途径。虽然有几个平台可供选择，但最近许多针对序列特异性靶向工具的进展都集中在开发基于成簇规律间隔短回文重复序列 - CRISPR 相关（CRISPR-Cas）的系统上。该系统使用无催化活性的 Cas 蛋白（dCas），可以作为不同模块化催化结构域（效应结构域）的载体，以控制基因的表达或改变表观遗传标记，如 DNA 甲基化。开发 CRISPR-dCas 系统的最新趋势包括创建可以将多个效应结构域靶向到单个位点的版本，靶向表观遗传变化，在某些情况下，在没有靶向构建体的情况下，可以遗传到下一代，以及将效应结构域和靶向策略结合起来，以产生协同作用，提高系统的功能或效率。本文综述并比较了 DNA 靶向技术、用于靶向转录控制和 epi 诱变的效应结构域，以及植物中使用的不同 CRISPR-dCas 系统。
