Universal and high-fidelity DNA single nucleotide polymorphism detection based on a CRISPR/Cas12a biochip

Chen，Yong1; Mei，Yixin1; Jiang，Xingyu1,2

2021-03-28

10.1039/d0sc05717g

Chemical Science   影响因子和分区

2041-6520

2041-6539

Single nucleotide polymorphisms (SNPs) are associated with many human diseases, so accurate and efficient SNP detection is of great significance for early diagnosis and clinical prognosis. This report proposes a universal and high-fidelity genotyping method in microfluidic point-of-care equipment based on the clustered regularly interspaced short palindromic repeat (CRISPR) system. Briefly, by systematically inserting the protospacer-adjacent-motif (PAM) sequence, we improved the universality of the CRISPR/Cas12a based SNP detection; by removing the complementary ssDNA and introducing an additional nucleotide mismatch, we improved the sensitivity and specificity. We preloaded the CRISPR/Cas12a reagents into the point-of-care biochip for automating the process, increasing the stability and long-term storage. This biochip enables us to rapidly and conveniently detect the genotypes within 20 min. In a practical application, the CRISPR/Cas12a biochip successfully distinguished three genotypes (homozygous wild type; the homozygous mutant type; and the heterozygous mutant type) of theCYP1A1*2(A4889G, rs1048943),CYP2C19*2(G681A, rs4244285),CYP2C9*3(A1075C, rs1057910), andCYP2C19*3(G636A, rs4986893) genes related to multiple cancers from 17 clinical blood samples. This CRISPR/Cas12a-based SNP genotyping method, being universal, accurate, and sensitive, will have broad applications in molecular diagnostics and clinical research.

SCI ; EI

英语

NI期刊 ; NI论文

第一 ; 通讯

WOS:000635768300024

20211410179846

Associative storage ; Clinical research ; Diagnosis ; Nucleotides ; Polymorphism

Bioengineering and Biology:461 ; Data Storage, Equipment and Techniques:722.1 ; Materials Science:951

2-s2.0-85103513599

Scopus

1.Department of Biomedical Engineering,Southern University of Science and Technology,Shenzhen,No. 1088, Xueyuan Rd., Xili, Nanshan District,China
2.Shenzhen Bay Laboratory,Shenzhen,518055,China

Chen，Yong,Mei，Yixin,Jiang，Xingyu. Universal and high-fidelity DNA single nucleotide polymorphism detection based on a CRISPR/Cas12a biochip[J]. Chemical Science,2021,12(12):4455-4462.

Chen，Yong,Mei，Yixin,&Jiang，Xingyu.(2021).Universal and high-fidelity DNA single nucleotide polymorphism detection based on a CRISPR/Cas12a biochip.Chemical Science,12(12),4455-4462.

Chen，Yong,et al."Universal and high-fidelity DNA single nucleotide polymorphism detection based on a CRISPR/Cas12a biochip".Chemical Science 12.12(2021):4455-4462.