Detection of the SARS-CoV-2 D614G mutation using engineered Cas12a guide RNA

文献类型： 外文期刊

第一作者： Meng, Qingzhou

作者： Meng, Qingzhou;Wang, Xinjie;Wang, Yanqun;Liu, Ming;Huang, Xingxu;Ma, Peixiang;Meng, Qingzhou;Dang, Lu;Wang, Xinjie;Wang, Xian;Yang, Guang;Ma, Peixiang;Wang, Xinjie;Ma, Xiaodong;Liu, Xinyi;Chi, Tian;Wang, Xian;Huang, Xingxu;Zhao, Qin

作者机构：

关键词： COVID-19; CRISPR; Cas12a; CRISPR-based detection; D614G; SARS-CoV-2

期刊名称：BIOTECHNOLOGY JOURNAL （ 影响因子：4.677； 五年影响因子：4.596 ）

ISSN： 1860-6768

年卷期：

页码：

收录情况： SCI

摘要： Detection of pathogens with single-nucleotide variations is indispensable for the disease tracing, but remains technically challenging. The D614G mutation in the SARS-CoV-2 spike protein is known to markedly enhance viral infectivity but is difficult to detect. Here, we report an effective approach called "synthetic mismatch integrated crRNA guided Cas12a detection" (symRNA-Cas12a) to detect the D614 and G614 variants effectively. Using this method, we systemically screened a pool of crRNAs that contain all the possible nucleotide substitutions covering the -2 to +2 positions around the mutation and identify one crRNA that can efficiently increase the detection specificity by 13-fold over the ancestral crRNA. With this selected crRNA, the symRNA-Cas12a assay can detect as low as 10 copies of synthetic mutant RNA and the results are confirmed to be accurate by Sanger sequencing. Overall, we have developed the symRNA-Cas12a method to specifically, sensitively and rapidly detect the SARS-CoV-2 D614G mutation.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序