Research progress of the CRISPR-Cas system in the detecting pathogen nucleic acids

DU Yao; GAO Hongdan; LIU Jiakun; LIU Xiaorong; XING Zhihao; ZHANG Tao; MA Dongli

doi:10.12211/2096-8280.2022-068

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Research progress of the CRISPR-Cas system in the detecting pathogen nucleic acids

Invited Review | 更新时间：2025-03-20

- Research progress of the CRISPR-Cas system in the detecting pathogen nucleic acids
- Synthetic Biology Journal Vol. 5, Issue 1, Pages: 202-216(2024)
- 作者机构：
  
  1.蚌埠医学院检验医学院，安徽蚌埠 233030
  2.深圳市儿童医院儿科研究所，广东深圳 518034
  3.中国科学院深圳先进技术研究所深圳合成生物研究所，广东深圳 518055
  4.深圳市海微生物科技有限公司，广东深圳 518057
  5.蚌埠医学院病原生物学教研室，安徽蚌埠 233030
- 作者简介：
- 基金信息：
- DOI：10.12211/2096-8280.2022-068
  CLC： R446.5
- Received：26 November 2022，
  
  Revised：2023-02-15，
  
  Published：29 February 2024
- 稿件说明：
移动端阅览
杜瑶，高宏丹，刘家坤，刘孝荣，邢志浩，张涛，马东礼. CRISPR-Cas系统在病原核酸检测中的研究进展［J］. 合成生物学， 2024， 5（1）： 202-216

DU Yao， GAO Hongdan， LIU Jiakun， LIU Xiaorong， XING Zhihao， ZHANG Tao， MA Dongli. Research progress of the CRISPR-Cas system in the detecting pathogen nucleic acids［J］. Synthetic Biology Journal， 2024， 5（1）： 202-216
杜瑶，高宏丹，刘家坤，刘孝荣，邢志浩，张涛，马东礼. CRISPR-Cas系统在病原核酸检测中的研究进展［J］. 合成生物学， 2024， 5（1）： 202-216 DOI： 10.12211/2096-8280.2022-068.

DU Yao， GAO Hongdan， LIU Jiakun， LIU Xiaorong， XING Zhihao， ZHANG Tao， MA Dongli. Research progress of the CRISPR-Cas system in the detecting pathogen nucleic acids［J］. Synthetic Biology Journal， 2024， 5（1）： 202-216 DOI： 10.12211/2096-8280.2022-068.

摘要

CRISPR-Cas系统作为原核生物获得性免疫系统，由簇状规则间隔短回文重复序列（clustered regularly interspaced short palindromic repeats， CRISPR）和CRISPR相关蛋白（CRISPR-associated proteins， Cas）构成，因其识别和切割特定DNA或RNA序列，而成为分子诊断领域研究的热点。研究人员利用Cas蛋白（Cas12、Cas13、Cas14、Cas3等）结合信号放大和转化技术（荧光法、电位法、比色法、侧向流动技术等），开发了许多高灵敏度、高特异性、低成本的诊断平台，为病原核酸检测提供了新途径。本文介绍了CRISPR-Cas系统的生物学机制及分类，总结现有的基于Cas蛋白反式切割活性开发的病原核酸检测技术，描述其特点、功能和应用场景，并对该系统的未来应用前景进行展望，期望CRISPR-Cas系统成为包括核酸检测在内的多靶标的理想检测平台。

Abstract

The CRISPR-Cas system consists of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins

which has become the focus of molecular diagnosis because it recognizes and cleaves specific DNA or RNA sequences. Using Cas proteins (Cas12

Cas13

Cas14

Cas3

etc.) combined with signal amplification and transformation techniques (fluorescence

potentiometric

colorimetric

lateral flow assay

etc.)

researchers have developed many diagnostic platforms with high sensitivity

good specificity

and low cost

which provide a new tool for detecting pathogen nucleic acids. This review presents the biological mechanism and classification of the CRISPR-Cas system

and also summarizes existing technologies for detecting pathogenic nucleic acids based on the trans-cleavage activity of Cas proteins

commenting their properties

functions and application scenarios

with future applications prospected based on the functional characteristics of the CRISPR-Cas system

which is expected to become an ideal detection platform for other multiple targets.

关键词

Keywords

references

HWANG H , HWANG B Y , BUENO J . Biomarkers in infectious diseases [J ] . Disease Markers , 2018 , 2018 : 8509127 .

库婷婷 , 刘倩 , 桑楠 , 等 . 几种典型呼吸道病毒的病原学特征及其检测方法 [J ] . 环境化学 , 2020 , 39 ( 4 ): 841 - 851 .

KU T T , LIU Q , SANG N , et al . Pathogenic characteristics and detection methods of several typical respiratory viruses [J ] . Environmental Chemistry , 2020 , 39 ( 4 ): 841 - 851 .

MATTHIJS G , SOUCHE E , ALDERS M , et al . Guidelines for diagnostic next-generation sequencing [J ] . European Journal of Human Genetics , 2016 , 24 ( 1 ): 2 - 5 .

MORENS D M , FAUCI A S . Emerging pandemic diseases: how we got to COVID-19 [J ] . Cell , 2020 , 182 ( 5 ): 1077 - 1092 .

张礼堃 , 邹秉杰 , 宋沁馨 , 等 . PCR技术在新冠病毒核酸检测中的应用 [J ] . 医学研究生学报 , 2021 , 34 ( 5 ): 539 - 544 .

ZHANG L K , ZOU B J , SONG Q X , et al . Application of PCR in novel coronavirus nucleic acid detection [J ] . Journal of Medical Postgraduates , 2021 , 34 ( 5 ): 539 - 544 .

PICKAR-OLIVER A , GERSBACH C A . The next generation of CRISPR-Cas technologies and applications [J ] . Nature Reviews Molecular Cell Biology , 2019 , 20 ( 8 ): 490 - 507 .

BHATTACHARYYA R P , THAKKU S G , HUNG D T . Harnessing CRISPR effectors for infectious disease diagnostics [J ] . ACS Infectious Diseases , 2018 , 4 ( 9 ): 1278 - 1282 .

ISHINO Y , SHINAGAWA H , MAKINO K , et al . Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli , and identification of the gene product [J ] . Journal of Bacteriology , 1987 , 169 ( 12 ): 5429 - 5433 .

JANSEN R , EMBDEN J D , GAASTRA W , et al . Identification of genes that are associated with DNA repeats in prokaryotes [J ] . Molecular Microbiology , 2002 , 43 ( 6 ): 1565 - 1575 .

BARRANGOU R . CRISPR-Cas systems and RNA-guided interference [J ] . Wiley Interdisciplinary Reviews RNA , 2013 , 4 ( 3 ): 267 - 278 .

GARNEAU J E , DUPUIS M È , VILLION M , et al . The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA [J ] . Nature , 2010 , 468 ( 7320 ): 67 - 71 .

YAO R L , LIU D , JIA X , et al . CRISPR-Cas9/Cas12a biotechnology and application in bacteria [J ] . Synthetic and Systems Biotechnology , 2018 , 3 ( 3 ): 135 - 149 .

JACKSON S A , MCKENZIE R E , FAGERLUND R D , et al . CRISPR-Cas: adapting to change [J ] . Science , 2017 , 356 ( 6333 ): eaal5056 .

HILLE F , RICHTER H , WONG S P , et al . The biology of CRISPR-Cas: backward and forward [J ] . Cell , 2018 , 172 ( 6 ): 1239 - 1259 .

GASIUNAS G , SINKUNAS T , SIKSNYS V . Molecular mechanisms of CRISPR-mediated microbial immunity [J ] . Cellular and Molecular Life Sciences , 2014 , 71 ( 3 ): 449 - 465 .

MAKAROVA K S , WOLF Y I , ALKHNBASHI O S , et al . An updated evolutionary classification of CRISPR-Cas systems [J ] . Nature Reviews Microbiology , 2015 , 13 ( 11 ): 722 - 736 .

MAKAROVA K S , WOLF Y I , IRANZO J , et al . Evolutionary classification of CRISPR-Cas systems: a burst of class 2 and derived variants [J ] . Nature Reviews Microbiology , 2020 , 18 ( 2 ): 67 - 83 .

MAKAROVA K S , ZHANG F , KOONIN E V . SnapShot: class 1 CRISPR-cas systems [J ] . Cell , 2017 , 168 ( 5 ): 946 - 946.e1 .

SHMAKOV S , ABUDAYYEH O O , MAKAROVA K S , et al . Discovery and functional characterization of diverse class 2 CRISPR-Cas systems [J ] . Molecular Cell , 2015 , 60 ( 3 ): 385 - 397 .

LI P P , WANG L , YANG J N , et al . Applications of the CRISPR-Cas system for infectious disease diagnostics [J ] . Expert Review of Molecular Diagnostics , 2021 , 21 ( 7 ): 723 - 732 .

BROUNS S J J , JORE M M , LUNDGREN M , et al . Small CRISPR RNAs guide antiviral defense in prokaryotes [J ] . Science , 2008 , 321 ( 5891 ): 960 - 964 .

HAYES R P , XIAO Y B , DING F , et al . Structural basis for promiscuous PAM recognition in type Ⅰ-E Cascade from E. coli [J ] . Nature , 2016 , 530 ( 7591 ): 499 - 503 .

ZHENG Y L , LI J , WANG B Y , et al . Endogenous typeⅠCRISPR-Cas: from foreign DNA defense to prokaryotic engineering [J ] . Frontiers in Bioengineering and Biotechnology , 2020 , 8 : 62 .

YOSHIMI K , TAKESHITA K , YAMAYOSHI S , et al . CRISPR-Cas3-based diagnostics for SARS-CoV-2 and influenza virus [J ] . iScience , 2022 , 25 ( 2 ): 103830 .

HOCHSTRASSER M L , TAYLOR D W , BHAT P , et al . CasA mediates Cas3-catalyzed target degradation during CRISPR RNA-guided interference [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2014 , 111 ( 18 ): 6618 - 6623 .

ZHAO H T , SHENG G , WANG J Y , et al . Crystal structure of the RNA-guided immune surveillance Cascade complex in Escherichia coli [J ] . Nature , 2014 , 515 ( 7525 ): 147 - 150 .

KAZLAUSKIENE M , KOSTIUK G , VENCLOVAS Č , et al . A cyclic oligonucleotide signaling pathway in typeⅢCRISPR-Cas systems [J ] . Science , 2017 , 357 ( 6351 ): 605 - 609 .

PINILLA-REDONDO R , MAYO-MUÑOZ D , RUSSEL J , et al . Type Ⅳ CRISPR-Cas systems are highly diverse and involved in competition between plasmids [J ] . Nucleic Acids Research , 2020 , 48 ( 4 ): 2000 - 2012 .

SHMAKOV S , SMARGON A , SCOTT D , et al . Diversity and evolution of class 2 CRISPR-Cas systems [J ] . Nature Reviews Microbiology , 2017 , 15 ( 3 ): 169 - 182 .

JIANG W Y , BIKARD D , COX D , et al . RNA-guided editing of bacterial genomes using CRISPR-Cas systems [J ] . Nature Biotechnology , 2013 , 31 ( 3 ): 233 - 239 .

JINEK M , CHYLINSKI K , FONFARA I , et al . A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity [J ] . Science , 2012 , 337 ( 6096 ): 816 - 821 .

SWARTS D C , JINEK M . Mechanistic insights into the cis - and trans -acting DNase activities of Cas12a [J ] . Molecular Cell , 2019 , 73 ( 3 ): 589 - 600.e4 .

ZENG R J , XU J H , LU L L , et al . Photoelectrochemical bioanalysis of microRNA on yolk-in-shell Au@CdS based on the catalytic hairpin assembly-mediated CRISPR-Cas12a system [J ] . Chemical Communications , 2022 , 58 ( 54 ): 7562 - 7565 .

CHEN H , LI Z Y , CHEN J S , et al . CRISPR/Cas12a-based electrochemical biosensor for highly sensitive detection of cTnI [J ] . Bioelectrochemistry , 2022 , 146 : 108167 .

KIM D , KIM J , HUR J K , et al . Erratum: Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells [J ] . Nature Biotechnology , 2016 , 34 ( 8 ): 888 .

XIN C C , YIN J H , YUAN S P , et al . Comprehensive assessment of miniature CRISPR-Cas12f nucleases for gene disruption [J ] . Nature Communications , 2022 , 13 : 5623 .

CASTELLE C J , WRIGHTON K C , THOMAS B C , et al . Genomic expansion of domain Archaea highlights roles for organisms from new phyla in anaerobic carbon cycling [J ] . Current Biology , 2015 , 25 ( 6 ): 690 - 701 .

HARRINGTON L B , BURSTEIN D , CHEN J S , et al . Programmed DNA destruction by miniature CRISPR-Cas14 enzymes [J ] . Science , 2018 , 362 ( 6416 ): 839 - 842 .

KHAN M Z , HAIDER S , MANSOOR S , et al . Targeting plant ssDNA viruses with engineered miniature CRISPR-Cas14a [J ] . Trends in Biotechnology , 2019 , 37 ( 8 ): 800 - 804 .

ANANTHARAMAN V , MAKAROVA K S , BURROUGHS A M , et al . Comprehensive analysis of the HEPN superfamily: identification of novel roles in intra-genomic conflicts, defense, pathogenesis and RNA processing [J ] . Biology Direct , 2013 , 8 : 15 .

ABUDAYYEH O O , GOOTENBERG J S , ESSLETZBICHLER P , et al . RNA targeting with CRISPR-Cas13 [J ] . Nature , 2017 , 550 ( 7675 ): 280 - 284 .

COX D B T , GOOTENBERG J S , ABUDAYYEH O O , et al . RNA editing with CRISPR-Cas13 [J ] . Science , 2017 , 358 ( 6366 ): 1019 - 1027 .

MAKAROVA K S , WOLF Y I , KOONIN E V . Classification and nomenclature of CRISPR-Cas systems: where from here? [J ] . The CRISPR Journal , 2018 , 1 ( 5 ): 325 - 336 .

CONG L , RAN F A , COX D , et al . Multiplex genome engineering using CRISPR/Cas systems [J ] . Science , 2013 , 339 ( 6121 ): 819 - 823 .

李洋 , 申晓林 , 孙新晓 , 等 . CRISPR基因编辑技术在微生物合成生物学领域的研究进展 [J ] . 合成生物学 , 2021 , 2 ( 1 ): 106 - 120 .

LI Y , SHEN X L , SUN X X , et al . Advances of CRISPR gene editing in microbial synthetic biology [J ] . Synthetic Biology Journal , 2021 , 2 ( 1 ): 106 - 120 .

PARDEE K , GREEN A A , TAKAHASHI M K , et al . Rapid, low-cost detection of Zika virus using programmable biomolecular components [J ] . Cell , 2016 , 165 ( 5 ): 1255 - 1266 .

HILTON I B , D’IPPOLITO A M , VOCKLEY C M , et al . Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers [J ] . Nature Biotechnology , 2015 , 33 ( 5 ): 510 - 517 .

SLOMOVICS , PARDEEK , COLLINSJ J . Synthetic biology devices for in vitro and in vivo diagnostics [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2015 , 112 ( 47 ): 14429 - 14435 .

LU T K , BOWERS J , KOERIS M S . Advancing bacteriophage-based microbial diagnostics with synthetic biology [J ] . Trends in Biotechnology , 2013 , 31 ( 6 ): 325 - 327 .

QUAN J , LANGELIER C , KUCHTA A , et al . FLASH: a next-generation CRISPR diagnostic for multiplexed detection of antimicrobial resistance sequences [J ] . Nucleic Acids Research , 2019 , 47 ( 14 ): e83 .

ZHOU W H , HU L , YING L M , et al . A CRISPR-Cas9-triggered strand displacement amplification method for ultrasensitive DNA detection [J ] . Nature Communications , 2018 , 9 : 5012 .

TSOU J H , LENG Q X , JIANG F . A CRISPR test for detection of circulating nuclei acids [J ] . Translational Oncology , 2019 , 12 ( 12 ): 1566 - 1573 .

SUN Y Y , YU L , LIU C X , et al . One-tube SARS-CoV-2 detection platform based on RT-RPA and CRISPR/Cas12a [J ] . Journal of Translational Medicine , 2021 , 19 ( 1 ): 74 .

LI S Y , CHENG Q X , WANG J M , et al . CRISPR-Cas12a-assisted nucleic acid detection [J ] . Cell Discovery , 2018 , 4 : 20 .

LI L X , LI S Y , WU N , et al . HOLMESv2: a CRISPR-Cas12b-assisted platform for nucleic acid detection and DNA methylation quantitation [J ] . ACS Synthetic Biology , 2019 , 8 ( 10 ): 2228 - 2237 .

TENG F , GUO L , CUI T T , et al . CDetection: CRISPR-Cas12b-based DNA detection with sub-attomolar sensitivity and single-base specificity [J ] . Genome Biology , 2019 , 20 ( 1 ): 132 .

DAI Y F , SOMOZA R A , WANG L , et al . Exploring the trans-cleavage activity of CRISPR-Cas12a (cpf1) for the development of a universal electrochemical biosensor [J ] . Angewandte Chemie (International Ed in English) , 2019 , 58 ( 48 ): 17399 - 17405 .

WU C S , CHEN Z , LI C Z , et al . CRISPR-Cas12a-empowered electrochemical biosensor for rapid and ultrasensitive detection of SARS-CoV-2 delta variant [J ] . Nano-Micro Letters , 2022 , 14 ( 1 ): 159 .

NGUYEN L T , GURIJALA J , RANANAWARE S R , et al . CRISPR-ENHANCE: an enhanced nucleic acid detection platform using Cas12a [J ] . Methods , 2022 , 203 : 116 - 124 .

DING X , YIN K , LI Z Y , et al . All-in-one dual CRISPR-Cas12a (AIOD-CRISPR) assay: a case for rapid, ultrasensitive and visual detection of novel coronavirus SARS-CoV-2 and HIV virus [EB/OL ] . BioRxiv . ( 2020-03-21 ) [ 2022-11-26 ] . https://www.biorxiv.org/content/10.1101/2020.03.19.998724v1 https://www.biorxiv.org/content/10.1101/2020.03.19.998724v1 .

NOURI R , JIANG Y Q , TANG Z F , et al . Detection of SARS-CoV-2 with solid-state CRISPR-Cas12a-assisted nanopores [J ] . Nano Letters , 2021 , 21 ( 19 ): 8393 - 8400 .

SAM I K , CHEN Y Y , MA J , et al . TB-QUICK: CRISPR-Cas12b-assisted rapid and sensitive detection of Mycobacterium tuberculosis [J ] . Journal of Infection , 2021 , 83 ( 1 ): 54 - 60 .

JOUNG J , LADHA A , SAITO M , et al . Point-of-care testing for COVID-19 using SHERLOCK diagnostics [EB/OL ] . MedRxiv . ( 2020-05-08 ) [ 2022-11-26 ] . https://www.medrxiv.org/content/10.1101/2020.05.04.20091231v1 https://www.medrxiv.org/content/10.1101/2020.05.04.20091231v1 .

JOUNG J , LADHA A , SAITO M , et al . Detection of SARS-CoV-2 with SHERLOCK one-pot testing [J ] . The New England Journal of Medicine , 2020 , 383 ( 15 ): 1492 - 1494 .

LU S H , TONG X H , HAN Y , et al . Fast and sensitive detection of SARS-CoV-2 RNA using suboptimal protospacer adjacent motifs for Cas12a [J ] . Nature Biomedical Engineering , 2022 , 6 ( 3 ): 286 - 297 .

AQUINO-JARQUIN G . CRISPR-Cas14 is now part of the artillery for gene editing and molecular diagnostic [J ] . Nanomedicine: Nanotechnology, Biology, and Medicine , 2019 , 18 : 428 - 431 .

WEI Y D , TAO Z Z , WAN L , et al . Aptamer-based Cas14a1 biosensor for amplification-free live pathogenic detection [J ] . Biosensors & Bioelectronics , 2022 , 211 : 114282 .

KELLNER M J , KOOB J G , GOOTENBERG J S , et al . SHERLOCK: nucleic acid detection with CRISPR nucleases [J ] . Nature Protocols , 2019 , 14 ( 10 ): 2986 - 3012 .

ARIZTI-SANZ J , FREIJE C A , STANTON A C , et al . Streamlined inactivation, amplification, and Cas13-based detection of SARS-CoV-2 [J ] . Nature Communications , 2020 , 11 : 5921 .

LIU T Y , KNOTT G J , SMOCK D C J , et al . Accelerated RNA detection using tandem CRISPR nucleases [J ] . Nature Chemical Biology , 2021 , 17 ( 9 ): 982 - 988 .

ACKERMAN C M , MYHRVOLD C , THAKKU S G , et al . Massively multiplexed nucleic acid detection with Cas13 [J ] . Nature , 2020 , 582 ( 7811 ): 277 - 282 .

WELCH N L , ZHU M L , HUA C , et al . Multiplexed CRISPR-based microfluidic platform for clinical testing of respiratory viruses and identification of SARS-CoV-2 variants [J ] . Nature Medicine , 2022 , 28 ( 5 ): 1083 - 1094 .

GOOTENBERG J S , ABUDAYYEH O O , KELLNER M J , et al . Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6 [J ] . Science , 2018 , 360 ( 6387 ): 439 - 444 .

YAN W X , HUNNEWELL P , ALFONSE L E , et al . Functionally diverse typeⅤCRISPR-Cas systems [J ] . Science , 2019 , 363 ( 6422 ): 88 - 91 .

LEUNG R K K , CHENG Q X , WU Z L , et al . CRISPR-Cas12-based nucleic acids detection systems [J ] . Methods , 2022 , 203 : 276 - 281 .

NGUYEN L T , SMITH B M , JAIN P K . Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection [J ] . Nature Communications , 2020 , 11 : 4906 .

CHEN J S , MA E B , HARRINGTON L B , et al . CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity [J ] . Science , 2018 , 360 ( 6387 ): 436 - 439 .

BROUGHTON J P , DENG X D , YU G X , et al . CRISPR-Cas12-based detection of SARS-CoV-2 [J ] . Nature Biotechnology , 2020 , 38 ( 7 ): 870 - 874 .

STRECKER J , JONES S , KOOPAL B , et al . Engineering of CRISPR-Cas12b for human genome editing [J ] . Nature Communications , 2019 , 10 : 212 .

ZHANG M Y , LIU C Z , SHI Y , et al . Selective endpoint visualized detection of Vibrio parahaemolyticus with CRISPR/Cas12a assisted PCR using thermal cycler for on-site application [J ] . Talanta , 2020 , 214 : 120818 .

MUKAMA O , WU J H , LI , Z Y , et al . An ultrasensitive and specific point-of-care CRISPR/Cas12 based lateral flow biosensor for the rapid detection of nucleic acids [J ] . Biosensors & Bioelectronics , 2020 , 159 : 112143 .

XIAO X X , LIN Z Q , HUANG X H , et al . Rapid and sensitive detection of Vibrio vulnificus using CRISPR/Cas12a combined with a recombinase-aided amplification assay [J ] . Frontiers in Microbiology , 2021 , 12 : 767315 .

TENG F , CUI T T , FENG G H , et al . Repurposing CRISPR-Cas12b for mammalian genome engineering [J ] . Cell Discovery , 2018 , 4 : 63 .

LIU X , BU S J , FENG J Q , et al . Electrochemical biosensor for detecting pathogenic bacteria based on a hybridization chain reaction and CRISPR-Cas12a [J ] . Analytical and Bioanalytical Chemistry , 2022 , 414 ( 2 ): 1073 - 1080 .

LI F , YE Q H , CHEN M T , et al . An ultrasensitive CRISPR/Cas12a based electrochemical biosensor for Listeria monocyto genes detection [J ] . Biosensors & Bioelectronics , 2021 , 179 : 113073 .

NOURI R , JIANG Y Q , LIAN X L , et al . Sequence-specific recognition of HIV-1 DNA with solid-state CRISPR-Cas12a-assisted nanopores (SCAN) [J ] . ACS Sensors , 2020 , 5 ( 5 ): 1273 - 1280 .

GORBALENYA A E , BAKER S C , BARIC R S , et al . The species severe acute respiratory syndrome-related coronavirus: classifying 2019-nCoV and naming it SARS-CoV-2 [J ] . Nature Microbiology , 2020 , 5 ( 4 ): 536 - 544 .

RHEE S Y , CLUTTER D , FESSEL W J , et al . Trends in the molecular epidemiology and genetic mechanisms of transmitted human immunodeficiency virus type 1 drug resistance in a large US clinic population [J ] . Clinical Infectious Diseases , 2019 , 68 ( 2 ): 213 - 221 .

ABUDAYYEH O O , GOOTENBERG J S , KONERMANN S , et al . C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector [J ] . Science , 2016 , 353 ( 6299 ): aaf5573 .

HADIDI A . Next-generation sequencing and CRISPR/Cas13 editing in viroid research and molecular diagnostics [J ] . Viruses , 2019 , 11 ( 2 ): 120 .

GOOTENBERG J S , ABUDAYYEH O O , LEE J W , et al . Nucleic acid detection with CRISPR-Cas13a/C2c2 [J ] . Science , 2017 , 356 ( 6336 ): 438 - 442 .

LEE R A , PUIG H , NGUYEN P Q , et al . Ultrasensitive CRISPR-based diagnostic f or field-applicable detection of Plasmodium species in symptomatic and asymptomatic malaria [J ] . Proceedings of the National Academy of Sciences of the United States of America , 2020 , 117 ( 41 ): 25722 - 25731 .

MYHRVOLD C , FREIJE C A , GOOTENBERG J S , et al . Field-deployable viral diagnostics using CRISPR-Cas13 [J ] . Science , 2018 , 360 ( 6387 ): 444 - 448 .

BARNES K G , LACHENAUER A E , NITIDO A , et al . Deployable CRISPR-Cas13a diagnostic tools to detect and report Ebola and Lassa virus cases in real-time [J ] . Nature Communications , 2020 , 11 : 4131 .

GACH P C , IWAI K , KIM P W , et al . Droplet microfluidics for synthetic biology [J ] . Lab on a Chip , 2017 , 17 ( 20 ): 3388 - 3400 .

IWAI K , WEHRS M , GARBER M , et al . Scalable and automated CRISPR-based strain engineering using droplet microfluidics [J ] . Microsystems & Nanoengineering , 2022 , 8 : 31 .

CHEN Y , MEI Y X , ZHAO X H , et al . Reagents-loaded, automated assay that integrates recombinase-aided amplification and Cas12a nucleic acid detection for a point-of-care test [J ] . Analytical Chemistry , 2020 , 92 ( 21 ): 14846 - 14852 .

PATCHSUNG M , JANTARUG K , PATTAMA A , et al . Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA [J ] . Nature Biomedical Engineering , 2020 , 4 ( 12 ): 1140 - 1149 .

EIBERG H , TROELSEN J , NIELSEN M , et al . Blue eye color in humans may be caused by a perfectly associated founder mutation in a regulatory element located within the HERC2 gene inhibiting OCA2 expression [J ] . Human Genetics , 2008 , 123 ( 2 ): 177 - 187 .

KOONIN E V , MAKAROVA K S , ZHANG F . Diversity, classification and evolution of CRISPR-Cas systems [J ] . Current Opinion in Microbiology , 2017 , 37 : 67 - 78 .

XIAO Y B , LUO M , DOLAN A E , et al . Structure basis for RNA-guided DNA degradation by Cascade and Cas3 [J ] . Science , 2018 , 361 ( 6397 ): eaat0839 .

KHAN W A , BARNEY R E , TSONGALIS G J . CRISPR-Cas13 enzymology rapidly detects SARS-CoV-2 fragments in a clinical setting [J ] . Journal of Clinical Virology , 2021 , 145 : 105019 .

LIANG M D , LI Z L , WANG W S , et al . A CRISPR-Cas12a-derived biosensing platform for the highly sensitive detection of diverse small molecules [J ] . Nature Communications , 2019 , 10 : 3672 .

ARCHAKOV A I , ASEEV A L , BYKOV V A , et al . Challenges of the human proteome project: 10-year experience of the Russian consortium [J ] . Journal of Proteome Research , 2019 , 18 ( 12 ): 4206 - 4214 .

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