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BEAT:一个用于从 Sanger 测序定量碱基编辑的 Python 程序。

BEAT: A Python Program to Quantify Base Editing from Sanger Sequencing.

机构信息

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.

出版信息

CRISPR J. 2019 Aug;2(4):223-229. doi: 10.1089/crispr.2019.0017. Epub 2019 Jul 18.

Abstract

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.

摘要

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。

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