Department of Surgery, Davis Heart and Lung Research Institute, Biomedical Sciences Graduate Program, Biophysics Graduate Program, The Ohio State University Wexner Medical Center, Columbus, Ohio; and Binghamton University, Binghamton, New York.
Department of Computer Science, Binghamton University, Binghamton, New York.
CRISPR J. 2019 Aug;2(4):223-229. doi: 10.1089/crispr.2019.0017. Epub 2019 Jul 18.
Through fusing CRISPR-Cas9 nickases with cytidine or adenine deaminases, a new paradigm-shifting class of genome-editing technology, termed "base editors," has recently been developed. Base editors mediate highly efficient, targeted single-base conversion without introducing double-stranded breaks. Analysis of base editing outcomes typically relies on imprecise enzymatic mismatch cleavage assays, time-consuming single-colony sequencing, or expensive next-generation deep sequencing. To overcome these limitations, several groups have recently developed computer programs to measure base-editing efficiency from fluorescence-based Sanger sequencing data such as Edit deconvolution by inference of traces in R (EditR), TIDER, and ICE. These approaches have greatly simplified the quantitation of base-editing experiments. However, the current Sanger sequencing tools lack the capability of batch analysis and producing high-quality images for publication. Here, we provide a ase diting nalysis ool (BEAT) written in Python to analyze and quantify the base-editing events from Sanger sequencing data in a batch manner, which can also produce intuitive, publication-ready base-editing images.
通过将 CRISPR-Cas9 核酸酶与胞嘧啶或腺嘌呤脱氨酶融合,一种新的、具有颠覆性的基因组编辑技术——“碱基编辑器”最近被开发出来。碱基编辑器介导高效、靶向的单碱基转换,而不会引入双链断裂。碱基编辑结果的分析通常依赖于不精确的酶切错配分析、耗时的单克隆测序或昂贵的下一代深度测序。为了克服这些限制,最近有几个研究小组开发了计算机程序,从基于荧光的 Sanger 测序数据(如通过 R 推断痕迹的编辑去卷积(EditR)、TIDER 和 ICE)来测量碱基编辑效率。这些方法大大简化了碱基编辑实验的定量分析。然而,目前的 Sanger 测序工具缺乏批量分析和生成高质量图像以供出版的能力。在这里,我们提供了一个基于 Python 的碱基编辑分析工具(BEAT),可以以批处理的方式分析和量化 Sanger 测序数据中的碱基编辑事件,还可以生成直观的、可用于出版的碱基编辑图像。