TY - JOUR
T1 - Accurate Detection of Rare Mutant Alleles by Target Base-Specific Cleavage with the CRISPR/Cas9 System
AU - Lee, Dongin
AU - Lee, Ji Hyun
AU - Bang, Duhee
N1 - Publisher Copyright:
© 2021 American Chemical Society.
PY - 2021/6/18
Y1 - 2021/6/18
N2 - The detection of low-frequency somatic mutations enables early diagnosis of disease; however, base-substitution errors that arise during genomic library preparation and high-throughput sequencing can lead to false diagnostic information. To discriminate true genomic alterations from technical errors, we developed spCas9-assisted true variant labeling sequencing (CARVE-seq), which detects low-frequency mutant alleles with high accuracy. CARVE-seq utilizes single-base discrimination during spCas9 cleavage reactions to exclude technical errors. Ten single nucleotide variants that recurrently occur in tumors were assayed by CARVE-seq using 20 ng reference samples, and 100% positive predictive value and specificity was observed, which proved the highly accurate performance of CARVE-seq.
AB - The detection of low-frequency somatic mutations enables early diagnosis of disease; however, base-substitution errors that arise during genomic library preparation and high-throughput sequencing can lead to false diagnostic information. To discriminate true genomic alterations from technical errors, we developed spCas9-assisted true variant labeling sequencing (CARVE-seq), which detects low-frequency mutant alleles with high accuracy. CARVE-seq utilizes single-base discrimination during spCas9 cleavage reactions to exclude technical errors. Ten single nucleotide variants that recurrently occur in tumors were assayed by CARVE-seq using 20 ng reference samples, and 100% positive predictive value and specificity was observed, which proved the highly accurate performance of CARVE-seq.
KW - CRISPR/Cas9
KW - accurate mutant DNA detection
KW - disease diagnosis
KW - labeling true variants
KW - rare mutant allele
KW - single nucleotide variant (SNV)
UR - http://www.scopus.com/inward/record.url?scp=85108236961&partnerID=8YFLogxK
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U2 - 10.1021/acssynbio.1c00056
DO - 10.1021/acssynbio.1c00056
M3 - Article
C2 - 34009946
AN - SCOPUS:85108236961
SN - 2161-5063
VL - 10
SP - 1451
EP - 1464
JO - ACS Synthetic Biology
JF - ACS Synthetic Biology
IS - 6
ER -