Journal ArticleDOI
Rolling Circular Amplification (RCA)-Assisted CRISPR/Cas9 Cleavage (RACE) for Highly Specific Detection of Multiple Extracellular Vesicle MicroRNAs
Ruixuan Wang,Ruixuan Wang,Ruixuan Wang,Xianxian Zhao,Xiaohui Chen,Xiaopei Qiu,Xiaopei Qiu,Guangchao Qing,Hong Zhang,Liangliang Zhang,Xiaolin Hu,Zhuoqi He,Daidi Zhong,Ying Wang,Yang Luo +14 more
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TLDR
The high consistence between the proposed approach RCA-assisted CRISPR/Cas9 cleavage (RACE) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) in detecting EV-derived miRNAs abundance from both cultured cancer cells and clinical lung cancer patients validated its robustness, revealing its potentials in the screening, diagnosis and prognosis of various diseases.Abstract:
Multiplexed detection of extracellular vesicle (EV)-derived microRNAs (miRNAs) plays a critical role in facilitating disease diagnosis and prognosis evaluation. Herein, we developed a highly specific nucleic acid detection platform for simultaneous quantification of several EV-derived miRNAs in constant temperature by integrating the advantages of a clustered regularly interspaced short palindromic repeats/CRISPR associated nucleases (CRISPR/Cas) system and rolling circular amplification (RCA) techniques. Particularly, the proposed approach demonstrated single-base resolution attributed to the dual-specific recognition from both padlock probe-mediated ligation and protospacer adjacent motif (PAM)-triggered cleavage. The high consistency between the proposed approach RCA-assisted CRISPR/Cas9 cleavage (RACE) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) in detecting EV-derived miRNAs' abundance from both cultured cancer cells and clinical lung cancer patients validated its robustness, revealing its potentials in the screening, diagnosis, and prognosis of various diseases. In summary, RACE is a powerful tool for multiplexed, specific detection of nucleic acids in point-of-care diagnostics and field-deployable analysis.read more
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Journal ArticleDOI
Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities.
Jeanne Elisabeth van Dongen,Johanna Theodora Wilhelmina Berendsen,Renske D.M. Steenbergen,Rob M. F. Wolthuis,Jan C.T. Eijkel,Loes I. Segerink +5 more
TL;DR: This review covers the technical aspects of integrating CRISPR/Cas technology in miniaturized sensors for analysis on-site and discusses the challenges of point-of-careCRISPR sensing.
Journal ArticleDOI
CRISPR/Cas13a Powered Portable Electrochemiluminescence Chip for Ultrasensitive and Specific MiRNA Detection.
TL;DR: Analysis results of miRNA from tumor cells also demonstrate the PECL‐CRISPR platform holds a promising potential for molecular diagnosis.
Journal ArticleDOI
Exosomes as a new frontier of cancer liquid biopsy
TL;DR: In this paper , a comprehensive overview on the conventional and novel technologies for exosome isolation, characterization and content detection is provided, and the roles of exosomes serving as potential biomarkers in liquid biopsy for the diagnosis, treatment monitoring, and prognosis prediction of cancer are summarized.
Journal ArticleDOI
Exosomes as a new frontier of cancer liquid biopsy
TL;DR: In this paper , a comprehensive overview on the conventional and novel technologies for exosome isolation, characterization and content detection is provided, and the roles of exosomes serving as potential biomarkers in liquid biopsy for the diagnosis, treatment monitoring, and prognosis prediction of cancer are summarized.
Journal ArticleDOI
CRISPR/cas systems redefine nucleic acid detection: Principles and methods.
Meng Wang,Rui Zhang,Jinming Li +2 more
TL;DR: A detailed classification and comprehensive discussion of recent works that harness these CRISPR-based diagnostic tools from a new perspective are provided, and current challenges and future perspectives of CRISpr-based diagnostics are outlined.
References
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Journal ArticleDOI
Genome engineering using the CRISPR-Cas9 system
F. Ann Ran,Patrick D. Hsu,Jason Wright,Vineeta Agarwala,Vineeta Agarwala,David A. Scott,Feng Zhang +6 more
TL;DR: A set of tools for Cas9-mediated genome editing via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, as well as generation of modified cell lines for downstream functional studies are described.
Journal ArticleDOI
Circulating microRNAs as stable blood-based markers for cancer detection
Patrick S. Mitchell,Rachael K. Parkin,Evan M. Kroh,Brian R. Fritz,Brian R. Fritz,Stacia K. Wyman,Era L. Pogosova-Agadjanyan,Amelia Peterson,Jennifer Noteboom,Kathy O'Briant,April Allen,Daniel W. Lin,Daniel W. Lin,Daniel W. Lin,Nicole Urban,Charles W. Drescher,Beatrice S. Knudsen,Derek L. Stirewalt,Robert Gentleman,Robert L. Vessella,Robert L. Vessella,Peter S. Nelson,Daniel Martin,Daniel Martin,Muneesh Tewari +24 more
TL;DR: It is shown here that miRNAs are present in human plasma in a remarkably stable form that is protected from endogenous RNase activity and established the measurement of tumor-derived mi RNAs in serum or plasma as an important approach for the blood-based detection of human cancer.
Journal ArticleDOI
Real-time quantification of microRNAs by stem–loop RT–PCR
Caifu Chen,Dana Ridzon,Adam Broomer,Zhaohui Zhou,Danny H. Lee,Julie T. Nguyen,Maura Barbisin,Nan Lan Xu,Vikram R. Mahuvakar,Mark R. Andersen,Kaiqin Lao,Kenneth J. Livak,Karl J. Guegler +12 more
TL;DR: A novel microRNA quantification method has been developed using stem–loop RT followed by TaqMan PCR analysis, which enables fast, accurate and sensitive miRNA expression profiling and can identify and monitor potential biomarkers specific to tissues or diseases.
Journal ArticleDOI
Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases
Xi Chen,Yi Ba,Lijia Ma,Lijia Ma,Xing Cai,Yuan Yin,Kehui Wang,Jigang Guo,Yujing Zhang,Jiangning Chen,Xing Guo,Qibin Li,Qibin Li,Xiaoying Li,Wenjing Wang,Yan Zhang,Jin Wang,Xueyuan Jiang,Yang Xiang,Chen Xu,Pingping Zheng,Juanbin Zhang,Ruiqiang Li,Hongjie Zhang,Xiaobin Shang,Ting Gong,Guang Ning,Jun Wang,Jun Wang,Ke Zen,Junfeng Zhang,Chen-Yu Zhang +31 more
TL;DR: It is demonstrated that miRNAs are present in the serum and plasma of humans and other animals such as mice, rats, bovine fetuses, calves, and horses, and can serve as potential biomarkers for the detection of various cancers and other diseases.
Journal ArticleDOI
MicroRNA therapeutics: towards a new era for the management of cancer and other diseases
TL;DR: Recent advances in the understanding of miRNAs in cancer and in other diseases are described and the challenge of identifying the most efficacious therapeutic candidates is discussed and a perspective on achieving safe and targeted delivery of miRNA therapeutics is provided.
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