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

doi:10.1002/biot.202100040

	Detection of the SARS-CoV-2 D614G mutation using engineered Cas12a guide RNA
	Meng, Qingzhou 1,2; Wang, Xinjie1,3,4 ; Wang, Yanqun 1; Dang, Lu 2; Liu, Xinyi 5; Ma, Xiaodong 4; Chi, Tian6 ; Wang, Xian3,6 ; Zhao, Qin 7; Yang, Guang3 ; Liu, Ming 1; Huang, Xingxu1,6 ; Ma, Peixiang1,3
	2021-06
发表期刊	BIOTECHNOLOGY JOURNAL (IF:3.2[JCR-2023],4.4[5-Year])
ISSN	1860-6768
EISSN	1860-7314
卷号	16 期号:6
发表状态	正式接收
DOI	10.1002/biot.202100040
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.
关键词	COVID‐ 19 CRISPR Cas12a CRISPR‐ based detection D614G SARS‐ CoV‐ 2
收录类别	SCI ; EI ; SCIE
语种	英语
WOS研究方向	Biochemical Research Methods ; Biotechnology & Applied Microbiology
WOS类目	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS记录号	WOS:000626545400001
出版者	WILEY-V C H VERLAG GMBH
原始文献类型	Article; Early Access
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/126095
专题	免疫化学研究所_特聘教授组_功能筛选实验室生命科学与技术学院_PI研究组_池天组生命科学与技术学院_PI研究组_邹燕组生命科学与技术学院_PI研究组_黄行许组生命科学与技术学院_硕士生
通讯作者	Liu, Ming; Huang, Xingxu; Ma, Peixiang
作者单位	1.Guangzhou Med Univ, Natl Clin Res Ctr Resp Dis, Natl Ctr Resp Med,Guangzhou Inst Resp Hlth, Guangzhou Lab,State Key Lab Resp Dis,Affiliated H, Guangzhou 510120, Peoples R China; 2.Guangzhou Med Univ, Affiliated Canc Hosp & Inst, Guangzhou, Peoples R China; 3.ShanghaiTech Univ, Shanghai Inst Adv Immunochem Studies, Shanghai, Peoples R China; 4.South China Normal Univ, Ctr Studies Psychol Applicat, Inst Brain Res & Rehabil, Guangdong Key Lab Mental Hlth & Cognit Sci, Guangzhou, Peoples R China; 5.Chinese Acad Agr Sci, Agr Genom Inst Shenzhen, Shenzhen, Peoples R China; 6.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China; 7.Northwest A&F Univ, Coll Vet Med, Dept Prevent Vet Med, Yangling, Shaanxi, Peoples R China
通讯作者单位	生命科学与技术学院; 免疫化学研究所
推荐引用方式 GB/T 7714	Meng, Qingzhou,Wang, Xinjie,Wang, Yanqun,et al. Detection of the SARS-CoV-2 D614G mutation using engineered Cas12a guide RNA[J]. BIOTECHNOLOGY JOURNAL,2021,16(6).
APA	Meng, Qingzhou.,Wang, Xinjie.,Wang, Yanqun.,Dang, Lu.,Liu, Xinyi.,...&Ma, Peixiang.(2021).Detection of the SARS-CoV-2 D614G mutation using engineered Cas12a guide RNA.BIOTECHNOLOGY JOURNAL,16(6).
MLA	Meng, Qingzhou,et al."Detection of the SARS-CoV-2 D614G mutation using engineered Cas12a guide RNA".BIOTECHNOLOGY JOURNAL 16.6(2021).