CRISPR-Cas9-Cytidine deaminase fusion enzymes-termed "base editors"-allow targeted editing of genomic deoxycytidine to deoxythymidine (C:G→T:A) without the need for double-stranded break induction. Base editors represent a paradigm shift in gene editing technology due to their unprecedented efficiency to mediate targeted, single-base conversion. However, current analysis of base editing outcomes rely on methods that are either imprecise or expensive and time-consuming. To overcome these limitations, we developed a simple, cost-effective, and accurate program to measure base editing efficiency from fluorescence-based Sanger sequencing, termed "EditR." We provide EditR as a free online tool or downloadable desktop application requiring a single Sanger sequencing file and guide RNA sequence. EditR is more accurate than enzymatic assays, and provides added insight to the position, type, and efficiency of base editing. Furthermore, EditR is likely amenable to quantify base editing from the recently developed adenosine deaminase base editors that act on either DNA (adenosine deaminase base editors [ABEs]) or RNA (REPAIRs) (catalyzes A:T→G:C). Collectively, we demonstrate that EditR is a robust, inexpensive tool that will facilitate the broad application of base editing technology, thereby fostering further innovation in this burgeoning field.

中文翻译：

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EditR：一种从Sanger测序中量化基础编辑的方法。

CRISPR-Cas9-胞苷脱氨酶融合酶被称为“碱基编辑器”-允许将基因组脱氧胞苷有针对性地编辑为脱氧胸苷（C：G→T：A），而无需双链断裂诱导。基础编辑器代表了基因编辑技术的一种范式转变，这是因为它们以前所未有的效率来介导目标化的单碱基转化。但是，当前对基本编辑结果的分析依赖于不精确，昂贵或费时的方法。为了克服这些限制，我们开发了一个简单，经济高效且准确的程序来测量基于荧光的Sanger测序（称为“ EditR”）的碱基编辑效率。我们提供EditR作为免费的在线工具或可下载的桌面应用程序，需要一个Sanger测序文件并指导RNA序列。EditR比酶法测定更准确，并提供了对碱基编辑的位置，类型和效率的更多了解。此外，EditR可能适合从最近开发的作用于DNA（腺苷脱氨酶碱基编辑器[ABEs]）或RNA（REPAIRs）（催化A：T→G：C）的腺苷脱氨酶碱基编辑器量化碱基编辑。总的来说，我们证明EditR是一款功能强大，价格便宜的工具，将有助于基础编辑技术的广泛应用，从而促进这一新兴领域的进一步创新。EditR可能适合根据最近开发的作用于DNA（腺苷脱氨酶碱基编辑器[ABEs]）或RNA（REPAIRs）（催化A：T→G：C）的腺苷脱氨酶碱基编辑器来量化碱基编辑。总的来说，我们证明EditR是一款功能强大且价格便宜的工具，将有助于基础编辑技术的广泛应用，从而促进这一新兴领域的进一步创新。EditR可能适合从最近开发的作用于DNA（腺苷脱氨酶碱基编辑器[ABEs]）或RNA（REPAIRs）（催化A：T→G：C）的腺苷脱氨酶碱基编辑器量化碱基编辑。总的来说，我们证明EditR是一款功能强大且价格便宜的工具，将有助于基础编辑技术的广泛应用，从而促进这一新兴领域的进一步创新。