RNA interference (RNAi) is a useful technique for knocking down a protein of interest, allowing for the study for the function of a gene product. However, RNAi techniques are prone to off-target effects, such as non-specific knockdown of genes besides the protein of interest. An important control and companion to RNAi knockdown experiments is the rescue experiment, wherein the gene function is restored by expression of an RNAi-resistant construct of the protein of interest. Generating an RNAi-resistant construct of the protein of interest involves generating silent mutations within the coding sequence of the protein so that the resulting amino acid product is the same, but the protein mRNA is no longer a target for the RNAi. Here, Synonymous Mutation Generator, a Python-based web tool that takes an input DNA coding sequence and outputs a synonymous DNA coding sequence that is RNAi-resistant, is described. This web tool should be a useful resource for researchers cloning RNAi-resistant constructs. Synonymous Mutation Generator is easy to use and can be found at jong2.pythonanywhere.com, and the source code is available on GitHub.

中文翻译：

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同义突变发生器：用于设计RNAi抗性序列的网络工具

RNA干扰（RNAi）是敲低目标蛋白质的有用技术，可用于研究基因产物的功能。但是，RNAi技术容易产生脱靶效应，例如除目标蛋白外，基因的非特异性敲低。RNAi敲除实验的重要对照和伴侣是拯救实验，其中通过表达目的蛋白质的RNAi抗性构建体来恢复基因功能。产生目的蛋白的RNAi抗性构建体涉及在蛋白的编码序列内产生沉默突变，以使所得的氨基酸产物相同，但蛋白mRNA不再是RNAi的靶标。在这里，同义变异发生器 描述了一种基于Python的网络工具，该工具采用输入的DNA编码序列并输出耐RNAi的同义DNA编码序列。对于研究人员克隆抗RNAi的构建体，该网络工具应该是有用的资源。同义突变发生器易于使用，可以在jong2.pythonanywhere.com上找到，其源代码可在GitHub上找到。