Research Group of Traditional Food, Research Division of Strategic Food Technology, Korea Food Research Institute, Wanju-gun 55365, Republic of Korea.
Division of Polar Life Sciences, Korea Polar Research Institute, Incheon 21990, Republic of Korea.
J Biotechnol. 2021 Nov 20;341:163-167. doi: 10.1016/j.jbiotec.2021.09.019. Epub 2021 Oct 1.
Despite the advantages of CRISPR/Cas9 technology in the food industry, controversy over its off-target effects exists. We engineered an industrial Saccharomyces cerevisiae strain isolated from a Korean rice wine starter, Nuruk, using CRISPR/Cas9 to decrease ethyl carbamate (EC) formation. We disrupted the CAR1 gene encoding arginase, which plays a key role in EC formation. Subsequently, we compared the whole genome of the engineered strain to that of the wild type by analyzing heterozygous and homozygous mutations through variant calling. Homozygous mutations in the genome of the engineered strains were identified as the target mutations in CAR1 induced by CRISPR/Cas9, and no other off-target effects were observed. Our findings have critical implications for the use of CRISRP/Cas9 technology in yeasts in the food industry.
尽管 CRISPR/Cas9 技术在食品工业中有许多优势，但围绕其脱靶效应仍存在争议。我们使用 CRISPR/Cas9 技术对从韩国米酒 starter Nuruk 中分离出来的工业酿酒酵母进行工程改造，以降低氨基甲酸乙酯（EC）的形成。我们敲除了编码精氨酸酶的 CAR1 基因，该基因在 EC 形成中起着关键作用。随后，我们通过变异calling 分析杂合和纯合突变，比较了工程菌株和野生型的全基因组。工程菌株基因组中的纯合突变被鉴定为 CRISPR/Cas9 诱导的 CAR1 中的靶突变，没有观察到其他脱靶效应。我们的研究结果对于 CRISPR/Cas9 技术在食品工业中应用于酵母具有重要意义。
