PMID- 21240280
OWN - NLM
STAT- MEDLINE
DCOM- 20110309
LR  - 20211020
IS  - 1548-7105 (Electronic)
IS  - 1548-7091 (Print)
IS  - 1548-7091 (Linking)
VI  - 8
IP  - 2
DP  - 2011 Feb
TI  - A transgenic mouse for in vivo detection of endogenous labeled mRNA.
PG  - 165-70
LID - 10.1038/nmeth.1551 [doi]
AB  - Live-cell single mRNA imaging is a powerful tool but has been restricted in 
      higher eukaryotes to artificial cell lines and reporter genes. We describe an 
      approach that enables live-cell imaging of single endogenous labeled mRNA 
      molecules transcribed in primary mammalian cells and tissue. We generated a 
      knock-in mouse line with an MS2 binding site (MBS) cassette targeted to the 3' 
      untranslated region of the essential beta-actin gene. As beta-actin-MBS was 
      ubiquitously expressed, we could uniquely address endogenous mRNA regulation in 
      any tissue or cell type. We simultaneously followed transcription from the 
      beta-actin alleles in real time and observed transcriptional bursting in response to 
      serum stimulation with precise temporal resolution. We tracked single endogenous 
      labeled mRNA particles being transported in primary hippocampal neurons. The MBS 
      cassette also enabled high-sensitivity fluorescence in situ hybridization (FISH), 
      allowing detection and localization of single beta-actin mRNA molecules in various 
      mouse tissues.
FAU - Lionnet, Timothee
AU  - Lionnet T
AD  - Department of Anatomy and Structural Biology, Albert Einstein College of 
      Medicine, Bronx, New York, USA.
FAU - Czaplinski, Kevin
AU  - Czaplinski K
FAU - Darzacq, Xavier
AU  - Darzacq X
FAU - Shav-Tal, Yaron
AU  - Shav-Tal Y
FAU - Wells, Amber L
AU  - Wells AL
FAU - Chao, Jeffrey A
AU  - Chao JA
FAU - Park, Hye Yoon
AU  - Park HY
FAU - de Turris, Valeria
AU  - de Turris V
FAU - Lopez-Jones, Melissa
AU  - Lopez-Jones M
FAU - Singer, Robert H
AU  - Singer RH
LA  - eng
GR  - F32 GM087122/GM/NIGMS NIH HHS/United States
GR  - R01 EB002060-25/EB/NIBIB NIH HHS/United States
GR  - F32-GM087122/GM/NIGMS NIH HHS/United States
GR  - R01 GM086217-04/GM/NIGMS NIH HHS/United States
GR  - R01 GM084364-18/GM/NIGMS NIH HHS/United States
GR  - GM84364/GM/NIGMS NIH HHS/United States
GR  - R01 GM086217/GM/NIGMS NIH HHS/United States
GR  - EB2060/EB/NIBIB NIH HHS/United States
GR  - R01 GM084364/GM/NIGMS NIH HHS/United States
GR  - GM86217/GM/NIGMS NIH HHS/United States
GR  - R01 EB002060/EB/NIBIB NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20110116
PL  - United States
TA  - Nat Methods
JT  - Nature methods
JID - 101215604
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Animals
MH  - Cell Survival
MH  - Cells, Cultured
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Mice
MH  - Mice, Transgenic/*genetics
MH  - RNA, Messenger/*analysis/genetics
MH  - Transcription, Genetic
PMC - PMC3076588
MID - NIHMS257921
EDAT- 2011/01/18 06:00
MHDA- 2011/03/10 06:00
PMCR- 2011/08/01
CRDT- 2011/01/18 06:00
PHST- 2010/09/23 00:00 [received]
PHST- 2010/12/10 00:00 [accepted]
PHST- 2011/01/18 06:00 [entrez]
PHST- 2011/01/18 06:00 [pubmed]
PHST- 2011/03/10 06:00 [medline]
PHST- 2011/08/01 00:00 [pmc-release]
AID - nmeth.1551 [pii]
AID - 10.1038/nmeth.1551 [doi]
PST - ppublish
SO  - Nat Methods. 2011 Feb;8(2):165-70. doi: 10.1038/nmeth.1551. Epub 2011 Jan 16.