Gardiner Jason, Zhao Jenny M, Chaffin Kendall, Jacobsen Steven E
Department of Molecular, Cell, and Developmental Biology, University of California Los Angeles, CA 90095, USA.
Institute for Society and Genetics, University of California, Los Angeles, CA 90095, USA.
Epigenomes. 2020 Jun 12;4(2):9. doi: 10.3390/epigenomes4020009.
DNA methylation is an important epigenetic mark involved in gene regulation and silencing of transposable elements. The presence or absence of DNA methylation at specific sites can influence nearby gene expression and cause phenotypic changes that remain stable over generations. Recently, development of new technologies has enabled the targeted addition or removal of DNA methylation at specific sites of the genome. Of these new technologies, the targeting of the catalytic domain of DOMAINS REARRANGED METHYLTRANSFERASE 2 (ntDRM2cd) offers a promising tool for the addition of DNA methylation as it can directly methylate DNA. However, the methylation targeting efficiency of constructs using ntDRM2cd thus far has been relatively low. Previous studies have shown that the use of different promoters or terminators can greatly improve genome-editing efficiencies. In this study, we systematically survey a variety of promoter and terminator combinations to identify optimal combinations to use when targeting the addition of DNA methylation in .
DNA甲基化是一种重要的表观遗传标记，参与基因调控和转座元件的沉默。特定位点DNA甲基化的存在或缺失会影响附近基因的表达，并导致在多代中保持稳定的表型变化。最近，新技术的发展使得在基因组的特定位点靶向添加或去除DNA甲基化成为可能。在这些新技术中，靶向结构域重排甲基转移酶2（ntDRM2cd）的催化结构域为添加DNA甲基化提供了一种有前景的工具，因为它可以直接使DNA甲基化。然而，迄今为止，使用ntDRM2cd构建体的甲基化靶向效率相对较低。先前的研究表明，使用不同的启动子或终止子可以大大提高基因组编辑效率。在本研究中，我们系统地调查了多种启动子和终止子组合，以确定在靶向添加DNA甲基化时使用的最佳组合。
