PMID- 25082846
OWN - NLM
STAT- MEDLINE
DCOM- 20150610
LR  - 20140926
IS  - 1755-3245 (Electronic)
IS  - 0008-6363 (Linking)
VI  - 104
IP  - 1
DP  - 2014 Oct 1
TI  - Rapid and highly efficient inducible cardiac gene knockout in adult mice using 
      AAV-mediated expression of Cre recombinase.
PG  - 15-23
LID - 10.1093/cvr/cvu174 [doi]
AB  - AIMS: Inducible gene targeting in mice using the Cre/LoxP system has become a 
      valuable tool to analyse the roles of specific genes in the adult heart. However, 
      the commonly used Myh6-MerCreMer system requires time-consuming breeding 
      schedules and is potentially associated with cardiac side effects, which may 
      result in transient cardiac dysfunction. The aim of our study was to establish a 
      rapid and simple system for cardiac gene inactivation in conditional knockout 
      mice by gene transfer of a Cre recombinase gene using adeno-associated viral 
      vectors of serotype 9 (AAV9). METHODS AND RESULTS: AAV9 vectors expressing Cre 
      under the control of a human cardiac troponin T promoter (AAV-TnT-Cre) enabled a 
      highly efficient Cre/LoxP switching in cardiomyocytes 2 weeks after injection 
      into 5- to 6-week-old ROSA26-LacZ reporter mice. Recombination efficiency was at 
      least as high as observed with the Myh6-MerCreMer system. No adverse side effects 
      were detected upon application of AAV-TnT-Cre. As proof of principle, we studied 
      AAV-TnT-Cre in a conditional knockout model (Srf-flex1 mice) to deplete the 
      myocardium of the transcription factor serum response factor (SRF). Four weeks 
      after AAV-TnT-Cre injection, a strong decrease in the cardiac expression of SRF 
      mRNA and protein was observed. Furthermore, mice developed a severe cardiac 
      dysfunction with increased interstitial fibrosis in accordance with the central 
      role of SRF for the expression of contractile and calcium trafficking proteins in 
      the heart. CONCLUSIONS: AAV9-mediated expression of Cre is a promising approach 
      for rapid and efficient conditional cardiac gene knockout in adult mice.
CI  - Published on behalf of the European Society of Cardiology. All rights reserved. (c) 
      The Author 2014. For permissions please email: journals.permissions@oup.com.
FAU - Werfel, Stanislas
AU  - Werfel S
AD  - Department of Internal Medicine III, University of Heidelberg, Im Neuenheimer 
      Feld 410, Heidelberg 69120, Germany Institute for Pharmacology and Toxicology, 
      Technische Universitat Munchen, Biedersteiner Str. 29, Munich 80802, Germany DZHK 
      (German Centre for Cardiovascular Research), Partner Site Munich Heart Alliance, 
      Munich, Germany.
FAU - Jungmann, Andreas
AU  - Jungmann A
AD  - Department of Internal Medicine III, University of Heidelberg, Im Neuenheimer 
      Feld 410, Heidelberg 69120, Germany.
FAU - Lehmann, Lorenz
AU  - Lehmann L
AD  - Department of Internal Medicine III, University of Heidelberg, Im Neuenheimer 
      Feld 410, Heidelberg 69120, Germany DZHK (German Center for Cardiovascular 
      Research), Partner Site Heidelberg/Mannheim, Germany.
FAU - Ksienzyk, Jan
AU  - Ksienzyk J
AD  - Department of Internal Medicine III, University of Heidelberg, Im Neuenheimer 
      Feld 410, Heidelberg 69120, Germany.
FAU - Bekeredjian, Raffi
AU  - Bekeredjian R
AD  - Department of Internal Medicine III, University of Heidelberg, Im Neuenheimer 
      Feld 410, Heidelberg 69120, Germany DZHK (German Center for Cardiovascular 
      Research), Partner Site Heidelberg/Mannheim, Germany.
FAU - Kaya, Ziya
AU  - Kaya Z
AD  - Department of Internal Medicine III, University of Heidelberg, Im Neuenheimer 
      Feld 410, Heidelberg 69120, Germany DZHK (German Center for Cardiovascular 
      Research), Partner Site Heidelberg/Mannheim, Germany.
FAU - Leuchs, Barbara
AU  - Leuchs B
AD  - Applied Tumorvirology, German Cancer Research Center, Heidelberg, Germany.
FAU - Nordheim, Alfred
AU  - Nordheim A
AD  - Interfaculty Institute for Cell Biology, Department of Molecular Biology, 
      University of Tubingen, Tubingen, Germany.
FAU - Backs, Johannes
AU  - Backs J
AD  - Department of Internal Medicine III, University of Heidelberg, Im Neuenheimer 
      Feld 410, Heidelberg 69120, Germany DZHK (German Center for Cardiovascular 
      Research), Partner Site Heidelberg/Mannheim, Germany.
FAU - Engelhardt, Stefan
AU  - Engelhardt S
AD  - Institute for Pharmacology and Toxicology, Technische Universitat Munchen, 
      Biedersteiner Str. 29, Munich 80802, Germany DZHK (German Centre for 
      Cardiovascular Research), Partner Site Munich Heart Alliance, Munich, Germany.
FAU - Katus, Hugo A
AU  - Katus HA
AD  - Department of Internal Medicine III, University of Heidelberg, Im Neuenheimer 
      Feld 410, Heidelberg 69120, Germany DZHK (German Center for Cardiovascular 
      Research), Partner Site Heidelberg/Mannheim, Germany.
FAU - Muller, Oliver J
AU  - Muller OJ
AD  - Department of Internal Medicine III, University of Heidelberg, Im Neuenheimer 
      Feld 410, Heidelberg 69120, Germany DZHK (German Center for Cardiovascular 
      Research), Partner Site Heidelberg/Mannheim, Germany 
      oliver.mueller@med.uni-heidelberg.de o.mueller@dkfz-heidelberg.de.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140731
PL  - England
TA  - Cardiovasc Res
JT  - Cardiovascular research
JID - 0077427
RN  - 0 (Gt(ROSA)26Sor non-coding RNA, mouse)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Untranslated)
RN  - 0 (Serum Response Factor)
RN  - 0 (Troponin T)
RN  - EC 2.7.7.- (Cre recombinase)
RN  - EC 2.7.7.- (Integrases)
SB  - IM
CIN - Cardiovasc Res. 2014 Oct 1;104(1):3-4. PMID: 25187523
MH  - Animals
MH  - Cardiomyopathies/genetics/metabolism/pathology/physiopathology
MH  - Dependovirus/enzymology/*genetics
MH  - Down-Regulation
MH  - Fibrosis
MH  - *Gene Knockdown Techniques
MH  - Genotype
MH  - Integrases/biosynthesis/*genetics
MH  - Lac Operon
MH  - Male
MH  - Mice, Inbred C57BL
MH  - Mice, Knockout
MH  - Myocytes, Cardiac/*metabolism/pathology
MH  - Phenotype
MH  - Promoter Regions, Genetic
MH  - RNA, Messenger/metabolism
MH  - RNA, Untranslated/genetics
MH  - Serum Response Factor/genetics/metabolism
MH  - Time Factors
MH  - Troponin T/genetics
MH  - Ventricular Dysfunction, Left/genetics/metabolism/pathology/physiopathology
MH  - Ventricular Function, Left
OTO - NOTNLM
OT  - Adeno-associated virus
OT  - Cardiomyopathy
OT  - Conditional transgenic mouse
OT  - Gene regulation
OT  - Serum response factor
EDAT- 2014/08/02 06:00
MHDA- 2015/06/11 06:00
CRDT- 2014/08/02 06:00
PHST- 2014/08/02 06:00 [entrez]
PHST- 2014/08/02 06:00 [pubmed]
PHST- 2015/06/11 06:00 [medline]
AID - cvu174 [pii]
AID - 10.1093/cvr/cvu174 [doi]
PST - ppublish
SO  - Cardiovasc Res. 2014 Oct 1;104(1):15-23. doi: 10.1093/cvr/cvu174. Epub 2014 Jul 
      31.