Biosensing bacterial 16S rDNA by microchip electrophoresis combined with a CRISPR system based on real-time crRNA/Cas12a formation.
Journal
RSC advances
ISSN: 2046-2069
Titre abrégé: RSC Adv
Pays: England
ID NLM: 101581657
Informations de publication
Date de publication:
04 Aug 2022
04 Aug 2022
Historique:
received:
16
05
2022
accepted:
26
07
2022
entrez:
31
8
2022
pubmed:
1
9
2022
medline:
1
9
2022
Statut:
epublish
Résumé
The accurate, simple and sensitive detection of bacterial infections at the early stage is highly valuable in preventing the spread of disease. Recently, CRISPR-Cas12a enzyme-derived nucleic acid detection methods have emerged along with the discovery of the indiscriminate single-stranded DNA (ssDNA) cleavage activity of Cas12a. These nucleic acid detection methods are made effective and sensitive by combining them with isothermal amplification technologies. However, most of the proposed CRISPR-Cas12a strategies involve Cas-crRNA complexes in the preassembled mode, which result in inevitable nonspecific background signals. Besides, the signal ssDNA used in these strategies needs tedious pre-labeling of the signal molecules. Herein, a post-assembly CRISPR-Cas12a method has been proposed based on target-induced transcription amplification and real-time crRNA generation for bacterial 16S rDNA biosensing. This strategy is label-free through the combination of microchip electrophoresis (MCE) detection. In addition, this method eliminates the need for a protospacer adjacent motif (PAM) on the target sequences, and has the potential to be an effective and simple method for nucleic acid detection and infectious disease diagnosis.
Identifiants
pubmed: 36043114
doi: 10.1039/d2ra03069a
pii: d2ra03069a
pmc: PMC9364175
doi:
Types de publication
Journal Article
Langues
eng
Pagination
22219-22225Informations de copyright
This journal is © The Royal Society of Chemistry.
Déclaration de conflit d'intérêts
There are no conflicts to declare.
Références
ACS Sens. 2020 May 22;5(5):1427-1435
pubmed: 32337966
Trends Biotechnol. 2019 Jul;37(7):730-743
pubmed: 30654914
Nanotechnology. 2019 Jan 11;30(2):025501
pubmed: 30411709
Biosens Bioelectron. 2021 May 1;179:113042
pubmed: 33662816
Science. 2007 Mar 23;315(5819):1709-12
pubmed: 17379808
Science. 2018 Apr 27;360(6387):381-382
pubmed: 29700254
J Chromatogr A. 2020 Mar 29;1615:460734
pubmed: 31791592
Biosens Bioelectron. 2020 May 1;155:112100
pubmed: 32090878
Anal Chem. 2021 Feb 23;93(7):3551-3558
pubmed: 33570925
Anal Chem. 2019 Mar 5;91(5):3452-3458
pubmed: 30667212
Talanta. 2020 Aug 1;215:120897
pubmed: 32312442
ACS Sens. 2020 Jun 26;5(6):1615-1623
pubmed: 32375473
Lab Chip. 2019 Apr 9;19(8):1397-1405
pubmed: 30847458
Science. 2018 Apr 27;360(6387):436-439
pubmed: 29449511
Cell. 2015 Oct 22;163(3):759-71
pubmed: 26422227
Talanta. 2021 Jan 15;222:121686
pubmed: 33167290
ACS Sens. 2019 Apr 26;4(4):841-848
pubmed: 30908029
Chem Commun (Camb). 2016 Dec 22;53(2):455-458
pubmed: 27966686
ACS Sens. 2019 Aug 23;4(8):2131-2139
pubmed: 31366194
Talanta. 2018 Sep 1;187:365-369
pubmed: 29853060
Sens Actuators B Chem. 2020 Dec 15;325:128780
pubmed: 32843820
Chem Commun (Camb). 2021 Jan 7;57(2):247-250
pubmed: 33306075
Angew Chem Int Ed Engl. 2019 Nov 25;58(48):17399-17405
pubmed: 31568601
Anal Chem. 2019 Jun 18;91(12):7524-7530
pubmed: 31117398
Anal Chem. 2019 Dec 3;91(23):14960-14966
pubmed: 31682108
Biosens Bioelectron. 2019 Aug 1;138:111302
pubmed: 31112917
Anal Chem. 2020 May 5;92(9):6702-6708
pubmed: 32272843
Chem Sci. 2020 Jun 19;11(28):7362-7368
pubmed: 33133487
Chem Soc Rev. 2012 Apr 21;41(8):3193-209
pubmed: 22331210