PMID- 36027801
OWN - NLM
STAT- MEDLINE
DCOM- 20220923
LR  - 20220923
IS  - 1873-4235 (Electronic)
IS  - 0956-5663 (Linking)
VI  - 216
DP  - 2022 Nov 15
TI  - Direct visualization of living bacterial genotypes using CRISPR/Cas12a-circular 
      reporter nanoprobes.
PG  - 114641
LID - S0956-5663(22)00681-9 [pii]
LID - 10.1016/j.bios.2022.114641 [doi]
AB  - Bacterial genotyping is important for understanding the complex microbiota. 
      Although fluorescence in situ hybridization (FISH) has enabled bacterial 
      community identification with high spatial resolution, its unavoidable cell 
      fixation steps and signal generation by multi-probe stacking greatly limit its 
      application in living bacterial genotyping. Here, we designed 
      polyethyleneimine-encapsulated CRISPR/Cas12a-circular reporter nanoprobes 
      (CasCLR) for rapid and sensitive visualization of gene information in living 
      bacteria. We found that, nanoprobe-based sequential delivery of Cas12a/crRNA and 
      circular reporter into bacteria allowed single genomic loci to initiate 
      trans-cleavage activity of Cas12a, thereby cleaving CLR to generate amplified 
      fluorescent signals for imaging of target gene. Using CasCLR, we can sensitively 
      analyze the percentage of target bacteria in co-culture experiments and directly 
      detect pathogenic bacteria in uncultured mouse gut microbe. In addition, CasCLR 
      has the ability to sensitively analyze specific genotype of microbial communities 
      in vivo. This nanobiotechnology-based bacterial gene analysis is expected to 
      advance understanding of in vivo bacterial cytogenetic information.
CI  - Copyright (c) 2022. Published by Elsevier B.V.
FAU - Li, Yanan
AU  - Li Y
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China.
FAU - Pei, Yiran
AU  - Pei Y
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China.
FAU - Huang, Di
AU  - Huang D
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China.
FAU - Xu, Ru
AU  - Xu R
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China.
FAU - Wu, Yonghua
AU  - Wu Y
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China.
FAU - Li, Yaqiong
AU  - Li Y
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China.
FAU - Zhou, Ruixue
AU  - Zhou R
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China.
FAU - Wei, Yongbin
AU  - Wei Y
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China.
FAU - Tan, Xiao
AU  - Tan X
AD  - Department of Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry & 
      Chemical Biology, Tsinghua University, Beijing, 100084, China.
FAU - Gao, Hua
AU  - Gao H
AD  - Department of Pathogen Biology, School of Basic Medical Sciences, Zhengzhou 
      University, Zhengzhou, 450001, China.
FAU - Shi, Jinjin
AU  - Shi J
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China.
FAU - Zhang, Zhenzhong
AU  - Zhang Z
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China. 
      Electronic address: zhangzhenzhong@zzu.edu.cn.
FAU - Zhang, Kaixiang
AU  - Zhang K
AD  - School of Pharmaceutical Sciences, Key Laboratory of Targeting Therapy and 
      Diagnosis for Critical Diseases, Collaborative Innovation Center of New Drug 
      Research and Safety Evaluation, State Key Laboratory of Esophageal Cancer 
      Prevention & Treatment, Zhengzhou University, Zhengzhou, 450001, China. 
      Electronic address: zhangkx@zzu.edu.cn.
FAU - Li, Jinghong
AU  - Li J
AD  - Department of Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry & 
      Chemical Biology, Tsinghua University, Beijing, 100084, China. Electronic 
      address: jhli@mail.tsinghua.edu.cn.
LA  - eng
PT  - Journal Article
DEP - 20220819
PL  - England
TA  - Biosens Bioelectron
JT  - Biosensors & bioelectronics
JID - 9001289
RN  - 0 (Bacterial Proteins)
RN  - 0 (CRISPR-Associated Proteins)
RN  - 9002-98-6 (Polyethyleneimine)
SB  - IM
MH  - Animals
MH  - Bacteria/genetics/metabolism
MH  - Bacterial Proteins/genetics
MH  - *Biosensing Techniques
MH  - *CRISPR-Associated Proteins/genetics
MH  - CRISPR-Cas Systems/genetics
MH  - Genotype
MH  - In Situ Hybridization, Fluorescence
MH  - Mice
MH  - Polyethyleneimine
OTO - NOTNLM
OT  - Bacterial genotype
OT  - Bioassay
OT  - Biopolymer materials
OT  - CRISPR/Cas12a
OT  - In vivo imaging
COIS- Declaration of competing interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2022/08/27 06:00
MHDA- 2022/09/24 06:00
CRDT- 2022/08/26 18:28
PHST- 2022/07/05 00:00 [received]
PHST- 2022/08/06 00:00 [revised]
PHST- 2022/08/14 00:00 [accepted]
PHST- 2022/08/27 06:00 [pubmed]
PHST- 2022/09/24 06:00 [medline]
PHST- 2022/08/26 18:28 [entrez]
AID - S0956-5663(22)00681-9 [pii]
AID - 10.1016/j.bios.2022.114641 [doi]
PST - ppublish
SO  - Biosens Bioelectron. 2022 Nov 15;216:114641. doi: 10.1016/j.bios.2022.114641. 
      Epub 2022 Aug 19.