PMID- 25400545
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20141117
LR  - 20200930
IS  - 1662-5099 (Print)
IS  - 1662-5099 (Electronic)
IS  - 1662-5099 (Linking)
VI  - 7
DP  - 2014
TI  - A transgenic mouse line for collecting ribosome-bound mRNA using the tetracycline 
      transactivator system.
PG  - 82
LID - 10.3389/fnmol.2014.00082 [doi]
LID - 82
AB  - Acquiring the gene expression profiles of specific neuronal cell-types is 
      important for understanding their molecular identities. Genome-wide gene 
      expression profiles of genetically defined cell-types can be acquired by 
      collecting and sequencing mRNA that is bound to epitope-tagged ribosomes (TRAP; 
      translating ribosome affinity purification). Here, we introduce a transgenic 
      mouse model that combines the TRAP technique with the tetracycline transactivator 
      (tTA) system by expressing EGFP-tagged ribosomal protein L10a (EGFP-L10a) under 
      control of the tetracycline response element (tetO-TRAP). This allows both 
      spatial control of EGFP-L10a expression through cell-type specific tTA 
      expression, as well as temporal regulation by inhibiting transgene expression 
      through the administration of doxycycline. We show that crossing tetO-TRAP mice 
      with transgenic mice expressing tTA under the Camk2a promoter (Camk2a-tTA) 
      results in offspring with cell-type specific expression of EGFP-L10a in CA1 
      pyramidal neurons and medium spiny neurons in the striatum. 
      Co-immunoprecipitation confirmed that EGFP-L10a integrates into a functional 
      ribosomal complex. In addition, collection of ribosome-bound mRNA from the 
      hippocampus yielded the expected enrichment of genes expressed in CA1 pyramidal 
      neurons, as well as a depletion of genes expressed in other hippocampal 
      cell-types. Finally, we show that crossing tetO-TRAP mice with transgenic Fos-tTA 
      mice enables the expression of EGFP-L10a in CA1 pyramidal neurons that are 
      activated during a fear conditioning trial. The tetO-TRAP mouse can be combined 
      with other tTA mouse lines to enable gene expression profiling of a variety of 
      different cell-types.
FAU - Drane, Laurel
AU  - Drane L
AD  - Department of Neuroscience, School of Medicine, Tufts University Boston, MA, USA 
      ; Graduate Program in Neuroscience, Sackler School of Graduate Biomedical 
      Sciences, Tufts University Boston, MA, USA.
FAU - Ainsley, Joshua A
AU  - Ainsley JA
AD  - Department of Neuroscience, School of Medicine, Tufts University Boston, MA, USA.
FAU - Mayford, Mark R
AU  - Mayford MR
AD  - Department of Molecular and Cellular Neuroscience, The Scripps Research Institute 
      La Jolla, CA, USA.
FAU - Reijmers, Leon G
AU  - Reijmers LG
AD  - Department of Neuroscience, School of Medicine, Tufts University Boston, MA, USA.
LA  - eng
GR  - R01 MH057368/MH/NIMH NIH HHS/United States
GR  - R21 MH099794/MH/NIMH NIH HHS/United States
GR  - DP2 OD006446/OD/NIH HHS/United States
GR  - P30 NS047243/NS/NINDS NIH HHS/United States
GR  - T32 NS061764/NS/NINDS NIH HHS/United States
PT  - Journal Article
DEP - 20141029
PL  - Switzerland
TA  - Front Mol Neurosci
JT  - Frontiers in molecular neuroscience
JID - 101477914
PMC - PMC4212621
OTO - NOTNLM
OT  - CA1 pyramidal neuron
OT  - Camk2a
OT  - Fos
OT  - TRAP
OT  - tetracycline transactivator
EDAT- 2014/11/18 06:00
MHDA- 2014/11/18 06:01
PMCR- 2014/01/01
CRDT- 2014/11/18 06:00
PHST- 2014/08/15 00:00 [received]
PHST- 2014/10/08 00:00 [accepted]
PHST- 2014/11/18 06:00 [entrez]
PHST- 2014/11/18 06:00 [pubmed]
PHST- 2014/11/18 06:01 [medline]
PHST- 2014/01/01 00:00 [pmc-release]
AID - 10.3389/fnmol.2014.00082 [doi]
PST - epublish
SO  - Front Mol Neurosci. 2014 Oct 29;7:82. doi: 10.3389/fnmol.2014.00082. eCollection 
      2014.