TY - JOUR
T1 - Plasmonically Enhanced CRISPR/Cas13a-Based Bioassay for Amplification-Free Detection of Cancer-Associated RNA
AU - Liu, Lin
AU - Wang, Zheyu
AU - Wang, Yixuan
AU - Luan, Jingyi
AU - Morrissey, Jeremiah J.
AU - Naik, Rajesh R.
AU - Singamaneni, Srikanth
N1 - Funding Information:
L.L. and Z.W. contributed equally to this work. The authors acknowledge support from National Institutes of Health (R21CA236652, R01DE027098, and R01CA141521) and National Science Foundation (CBET‐1900277, CBET‐2027145). The authors also acknowledge support from the Air Force Research Laboratory. The authors thank Prof. Rohit Pappu and Dr. Matthew King for helping with Cas13a protein purification. The authors also thank Nano Research Facility (NRF) and Institute of Materials Science and Engineering (IMSE) at Washington University for providing access to electron microscopy facilities
Funding Information:
L.L. and Z.W. contributed equally to this work. The authors acknowledge support from National Institutes of Health (R21CA236652, R01DE027098, and R01CA141521) and National Science Foundation (CBET-1900277, CBET-2027145). The authors also acknowledge support from the Air Force Research Laboratory. The authors thank Prof. Rohit Pappu and Dr. Matthew King for helping with Cas13a protein purification. The authors also thank Nano Research Facility (NRF) and Institute of Materials Science and Engineering (IMSE) at Washington University for providing access to electron microscopy facilities
Publisher Copyright:
© 2021 Wiley-VCH GmbH
PY - 2021/10/20
Y1 - 2021/10/20
N2 - Novel methods that enable sensitive, accurate and rapid detection of RNA would not only benefit fundamental biological studies but also serve as diagnostic tools for various pathological conditions, including bacterial and viral infections and cancer. Although highly sensitive, existing methods for RNA detection involve long turn-around time and extensive capital equipment. Here, an ultrasensitive and amplification-free RNA quantification method is demonstrated by integrating CRISPR-Cas13a system with an ultrabright fluorescent nanolabel, plasmonic fluor. This plasmonically enhanced CRISPR-powered assay exhibits nearly 1000-fold lower limit-of-detection compared to conventional assay relying on enzymatic reporters. Using a xenograft tumor mouse model, it is demonstrated that this novel bioassay can be used for ultrasensitive and quantitative monitoring of cancer biomarker (lncRNA H19). The novel biodetection approach described here provides a rapid, ultrasensitive, and amplification-free strategy that can be broadly employed for detection of various RNA biomarkers, even in resource-limited settings.
AB - Novel methods that enable sensitive, accurate and rapid detection of RNA would not only benefit fundamental biological studies but also serve as diagnostic tools for various pathological conditions, including bacterial and viral infections and cancer. Although highly sensitive, existing methods for RNA detection involve long turn-around time and extensive capital equipment. Here, an ultrasensitive and amplification-free RNA quantification method is demonstrated by integrating CRISPR-Cas13a system with an ultrabright fluorescent nanolabel, plasmonic fluor. This plasmonically enhanced CRISPR-powered assay exhibits nearly 1000-fold lower limit-of-detection compared to conventional assay relying on enzymatic reporters. Using a xenograft tumor mouse model, it is demonstrated that this novel bioassay can be used for ultrasensitive and quantitative monitoring of cancer biomarker (lncRNA H19). The novel biodetection approach described here provides a rapid, ultrasensitive, and amplification-free strategy that can be broadly employed for detection of various RNA biomarkers, even in resource-limited settings.
KW - CRISPR-Cas13a
KW - RNA detection
KW - gold nanorods
KW - nanolabels
KW - plasmon-enhanced fluorescence
UR - http://www.scopus.com/inward/record.url?scp=85112607980&partnerID=8YFLogxK
U2 - 10.1002/adhm.202100956
DO - 10.1002/adhm.202100956
M3 - Article
C2 - 34369102
AN - SCOPUS:85112607980
SN - 2192-2640
VL - 10
JO - Advanced Healthcare Materials
JF - Advanced Healthcare Materials
IS - 20
M1 - 2100956
ER -