TY - JOUR
T1 - Digital CRISPR systems for the next generation of nucleic acid quantification
AU - Politza, Anthony J.
AU - Nouri, Reza
AU - Guan, Weihua
N1 - Funding Information:
This work was supported by the National Institutes of Health ( R61AI147419 ) and National Science Foundation ( 1902503 , 1912410 , 2045169 ). Any opinions, findings, and conclusions or recommendations expressed in this work are those of the authors and do not necessarily reflect the views of the National Science Foundation and National Institutes of Health.
Publisher Copyright:
© 2023 Elsevier B.V.
PY - 2023/2
Y1 - 2023/2
N2 - Digital CRISPR (dCRISPR) assays are an emerging platform of molecular diagnostics. Digital platforms introduce absolute quantification and increased sensitivity to bulk CRISPR assays. With ultra-specific targeting, isothermal operation, and rapid detection, dCRISPR systems are well-prepared to lead the field of molecular diagnostics. Here we summarized the common Cas proteins used in CRISPR detection assays. The methods of digital detection and critical performance factors are examined. We formed three strategies to frame the landscape of dCRISPR systems: (1) amplification free, (2) in-partition amplification, and (3) two-stage amplification. We also compared the performance of all systems through the limit of detection (LOD), testing time, and figure of merit (FOM). This work summarizes the details of digital CRISPR platforms to guide future development. We envision that improvements to LOD and dynamic range will position dCRISPR as the leading platform for the next generation of molecular biosensing.
AB - Digital CRISPR (dCRISPR) assays are an emerging platform of molecular diagnostics. Digital platforms introduce absolute quantification and increased sensitivity to bulk CRISPR assays. With ultra-specific targeting, isothermal operation, and rapid detection, dCRISPR systems are well-prepared to lead the field of molecular diagnostics. Here we summarized the common Cas proteins used in CRISPR detection assays. The methods of digital detection and critical performance factors are examined. We formed three strategies to frame the landscape of dCRISPR systems: (1) amplification free, (2) in-partition amplification, and (3) two-stage amplification. We also compared the performance of all systems through the limit of detection (LOD), testing time, and figure of merit (FOM). This work summarizes the details of digital CRISPR platforms to guide future development. We envision that improvements to LOD and dynamic range will position dCRISPR as the leading platform for the next generation of molecular biosensing.
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U2 - 10.1016/j.trac.2023.116917
DO - 10.1016/j.trac.2023.116917
M3 - Review article
C2 - 36744100
AN - SCOPUS:85146261218
SN - 0165-9936
VL - 159
JO - TrAC - Trends in Analytical Chemistry
JF - TrAC - Trends in Analytical Chemistry
M1 - 116917
ER -