PMID- 35149451
OWN - NLM
STAT- MEDLINE
DCOM- 20220315
LR  - 20220315
IS  - 1873-4235 (Electronic)
IS  - 0956-5663 (Linking)
VI  - 204
DP  - 2022 May 15
TI  - Live imaging of RNA and RNA splicing in mammalian cells via the 
      dcas13a-SunTag-BiFC system.
PG  - 114074
LID - S0956-5663(22)00114-2 [pii]
LID - 10.1016/j.bios.2022.114074 [doi]
AB  - Dynamic tracking of the localization of RNA molecules (nucleus and/or cytoplasm) 
      and RNA splicing in living cells plays an important role in understanding their 
      functions. However, a lack of dynamic imaging and high background fluorescence 
      have been reported in the fluorescence in situ hybridization (FISH). Here, we 
      developed a new tool, the dcas13a-SunTag-BiFC system, which fused the dLwacas13a 
      and SunTag systems. dLwacas13a is used as a tracker to target specific RNAs, 
      while SunTag recruits split Venus fluorescent proteins to label targeted RNAs. 
      Our results showed that 4 x NLS-dCas13a-24 x SunTag-BiFC and 2 x NLS- 
      dCas13a-24 x SunTag-BiFC systems can be used for imaging of endogenous RNA foci 
      in the nucleus (Xist) and cytoplasm (Ppib and stress granules) in living cells, 
      respectively. Compared to 12x MS2-MCP system, the dcas13a-SunTag-BiFC system 
      showed a better performance of mRNA foci tracking in live cells. Furthermore, we 
      confirmed the premature termination codon (PTC)-induced exon skipping of Oxt RNA 
      using the dcas13a-SunTag-BiFC and MS2-MCP systems in the nucleus. Thus, the 
      dcas13a-SunTag-BiFC system will facilitate the study of RNA localization in 
      living cells and provide new insights into RNA translocation and splicing.
CI  - Copyright (c) 2022 Elsevier B.V. All rights reserved.
FAU - Chen, Mao
AU  - Chen M
AD  - Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal 
      Science, Jilin University, Changchun, 130062, China.
FAU - Sui, Tingting
AU  - Sui T
AD  - Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal 
      Science, Jilin University, Changchun, 130062, China.
FAU - Yang, Li
AU  - Yang L
AD  - Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal 
      Science, Jilin University, Changchun, 130062, China.
FAU - Qian, Yuqiang
AU  - Qian Y
AD  - Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal 
      Science, Jilin University, Changchun, 130062, China.
FAU - Liu, Zhiquan
AU  - Liu Z
AD  - Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal 
      Science, Jilin University, Changchun, 130062, China.
FAU - Liu, Yongsai
AU  - Liu Y
AD  - Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal 
      Science, Jilin University, Changchun, 130062, China.
FAU - Wang, Gerong
AU  - Wang G
AD  - Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal 
      Science, Jilin University, Changchun, 130062, China.
FAU - Lai, Liangxue
AU  - Lai L
AD  - Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal 
      Science, Jilin University, Changchun, 130062, China; CAS Key Laboratory of 
      Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and 
      Regenerative Medicine, South China Institute for Stem Cell Biology and 
      Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese 
      Academy of Sciences, Guangzhou, 510530, China. Electronic address: 
      lai_liangxue@gibh.ac.cn.
FAU - Li, Zhanjun
AU  - Li Z
AD  - Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal 
      Science, Jilin University, Changchun, 130062, China. Electronic address: 
      lizj_1998@jlu.edu.cn.
LA  - eng
PT  - Journal Article
DEP - 20220204
PL  - England
TA  - Biosens Bioelectron
JT  - Biosensors & bioelectronics
JID - 9001289
RN  - 0 (RNA, Messenger)
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - Animals
MH  - *Biosensing Techniques
MH  - In Situ Hybridization, Fluorescence
MH  - *RNA/genetics/metabolism
MH  - RNA Splicing
MH  - RNA, Messenger/genetics
OTO - NOTNLM
OT  - Cas13
OT  - Live imaging
OT  - RNA
OT  - RNA splicing
EDAT- 2022/02/13 06:00
MHDA- 2022/03/16 06:00
CRDT- 2022/02/12 05:35
PHST- 2021/09/11 00:00 [received]
PHST- 2022/01/19 00:00 [revised]
PHST- 2022/02/02 00:00 [accepted]
PHST- 2022/02/13 06:00 [pubmed]
PHST- 2022/03/16 06:00 [medline]
PHST- 2022/02/12 05:35 [entrez]
AID - S0956-5663(22)00114-2 [pii]
AID - 10.1016/j.bios.2022.114074 [doi]
PST - ppublish
SO  - Biosens Bioelectron. 2022 May 15;204:114074. doi: 10.1016/j.bios.2022.114074. 
      Epub 2022 Feb 4.