Laboratory time scale evolution in vivo relies on the generation of large, mutationally diverse gene libraries to rapidly explore biomolecule sequence landscapes. Traditional global mutagenesis methods are problematic because they introduce many off-target mutations that are often lethal and can engender false positives. We report the development and application of the MutaT7 chimera, a potent and highly targeted in vivo mutagenesis agent. MutaT7 utilizes a DNA-damaging cytidine deaminase fused to a processive RNA polymerase to continuously direct mutations to specific, well-defined DNA regions of any relevant length. MutaT7 thus provides a mechanism for in vivo targeted mutagenesis across multi-kb DNA sequences. MutaT7 should prove useful in diverse organisms, opening the door to new types of in vivo evolution experiments.

中文翻译：

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一种进行性蛋白质嵌合体在体内引入跨定义 DNA 区域的突变

体内实验室时间尺度进化依赖于大型、突变多样的基因库的生成，以快速探索生物分子序列景观。传统的全局诱变方法存在问题，因为它们引入了许多脱靶突变，这些突变通常是致命的并且会产生误报。我们报告了 MutaT7 嵌合体的开发和应用，MutaT7 嵌合体是一种有效且高度靶向的体内诱变剂。MutaT7 利用与加工性 RNA 聚合酶融合的破坏 DNA 的胞苷脱氨酶，持续将突变定向到任何相关长度的特定、明确定义的 DNA 区域。因此，MutaT7 提供了一种跨多 kb DNA 序列的体内靶向诱变机制。MutaT7 应该在不同的生物体中被证明是有用的，为新型体内进化实验打开了大门。