PMID- 30731369
OWN - NLM
STAT- MEDLINE
DCOM- 20190812
LR  - 20190812
IS  - 1879-1506 (Electronic)
IS  - 0003-9969 (Linking)
VI  - 99
DP  - 2019 Mar
TI  - Deletion of cas3 gene in Streptococcus mutans affects biofilm formation and 
      increases fluoride sensitivity.
PG  - 190-197
LID - S0003-9969(18)30624-1 [pii]
LID - 10.1016/j.archoralbio.2019.01.016 [doi]
AB  - OBJECTIVE: The goal of this study was to analyze the impact of cas3 gene on the 
      biofilm formation and virulence gene expression in S. mutans, since our previous 
      studies have found a connection between CRISPR/Cas systems and biofilm formation 
      in S. mutans. METHODS: The cas3 gene in-frame deletion strains of S. mutans UA159 
      was constructed by a two-step transformation procedure and the cas3 mutant strain 
      was complemented in trans. The biofilm biomass was measured by crystal violet 
      staining, and the synthesis of exopolysaccharides (EPS) was measured by the 
      anthrone-sulfuric method. Biofilm analysis and structural imaging was using 
      confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) 
      assays. The fluorescence in situ hybridization (FISH) was used to analyze the 
      spatiotemporal interactions between S. mutans and Streptococcus sanguinis. 
      Fluoride sensitivity was determined using fluoride tolerance assays. The 
      expression of biofilm formation related genes was evaluated by qRT-PCR. RESULTS: 
      Our results showed that S. mutans cas3 deletion strain formed less biofilm and 
      became less competitive when it was co-cultured with S. sanguinis under fluoride 
      treatment. The expression levels of virulence genes including vicR, gtfC, smu0630 
      and comDE were significantly downregulated. CONCLUSIONS: The cas3 gene in S. 
      mutans could regulate biofilm formation and fluoride resistance, consequently 
      affecting S. mutans competitiveness in a dual-species biofilm model under 
      fluoride treatment. These results also provide a potential strategy for enhancing 
      fluoride specificity, with cas3 gene as a potential genetic target in the 
      modulation of oral microecology and the treatment of dental caries.
CI  - Copyright (c) 2019 Elsevier Ltd. All rights reserved.
FAU - Tang, Boyu
AU  - Tang B
AD  - State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral 
      Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
FAU - Gong, Tao
AU  - Gong T
AD  - State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral 
      Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
FAU - Zhou, Xuedong
AU  - Zhou X
AD  - State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral 
      Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
FAU - Lu, Miao
AU  - Lu M
AD  - State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral 
      Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
FAU - Zeng, Jumei
AU  - Zeng J
AD  - Department of Infectious Diseases, Boston Children's Hospital, Harvard Medical 
      School, Boston, MA, USA.
FAU - Peng, Xian
AU  - Peng X
AD  - State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral 
      Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
FAU - Wang, Shida
AU  - Wang S
AD  - State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral 
      Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
FAU - Li, Yuqing
AU  - Li Y
AD  - State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral 
      Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China. 
      Electronic address: liyuqing@scu.edu.cn.
LA  - eng
PT  - Journal Article
DEP - 20190129
PL  - England
TA  - Arch Oral Biol
JT  - Archives of oral biology
JID - 0116711
RN  - 0 (Bacterial Proteins)
RN  - 0 (CRISPR-Associated Proteins)
RN  - 0 (Escherichia coli Proteins)
RN  - 0 (Smu0630 protein, Streptococcus mutans)
RN  - 0 (YycF protein, Bacteria)
RN  - EC 3.6.4.- (DNA Helicases)
RN  - EC 3.6.4.12 (ygcB protein, E coli)
RN  - Q80VPU408O (Fluorides)
MH  - Bacterial Proteins/genetics
MH  - Biofilms/*growth & development
MH  - Biomass
MH  - CRISPR-Associated Proteins/*genetics
MH  - CRISPR-Cas Systems
MH  - Coculture Techniques
MH  - DNA Helicases
MH  - Dental Caries
MH  - Drug Resistance, Bacterial/genetics
MH  - Drug Tolerance
MH  - Escherichia coli Proteins
MH  - Fluorides/*pharmacology
MH  - Gene Deletion
MH  - Gene Expression Regulation, Bacterial
MH  - Genes, Bacterial/genetics
MH  - In Situ Hybridization, Fluorescence
MH  - Microbial Sensitivity Tests
MH  - Microscopy, Confocal
MH  - Microscopy, Electron, Scanning
MH  - Streptococcus mutans/*drug effects/*genetics/*metabolism
MH  - Streptococcus sanguis/physiology
MH  - Transcriptome
MH  - Virulence/genetics
OTO - NOTNLM
OT  - Biofilm formation
OT  - CRISPR
OT  - Fluoride resistance
OT  - Streptococcus mutans
OT  - cas3
EDAT- 2019/02/08 06:00
MHDA- 2019/08/14 06:00
CRDT- 2019/02/08 06:00
PHST- 2018/09/19 00:00 [received]
PHST- 2019/01/12 00:00 [revised]
PHST- 2019/01/26 00:00 [accepted]
PHST- 2019/02/08 06:00 [pubmed]
PHST- 2019/08/14 06:00 [medline]
PHST- 2019/02/08 06:00 [entrez]
AID - S0003-9969(18)30624-1 [pii]
AID - 10.1016/j.archoralbio.2019.01.016 [doi]
PST - ppublish
SO  - Arch Oral Biol. 2019 Mar;99:190-197. doi: 10.1016/j.archoralbio.2019.01.016. Epub 
      2019 Jan 29.