PMID- 22125074
OWN - NLM
STAT- MEDLINE
DCOM- 20120316
LR  - 20211203
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 821
DP  - 2012
TI  - Utilizing a retroviral RNAi system to investigate in vivo mTOR functions in T 
      cells.
PG  - 305-16
LID - 10.1007/978-1-61779-430-8_19 [doi]
AB  - RNA interference (RNAi) is an intracellular mechanism for silencing gene 
      expression utilizing short fragments of double-strand RNA that are complementary 
      to the target messenger RNA. This gene silencing technique has now become an 
      invaluable research tool due to its specific and strong repressive effect on a 
      target transcript. We have recently applied a retrovirus-based RNAi system to 
      investigate the in vivo role of the mammalian target of rapamycin (mTOR) in 
      antigen-specific CD8 T cells, and have found that mTOR regulates memory CD8 
      T-cell differentiation. Here, we provide a detailed protocol for knocking down 
      mTOR and its related molecules (raptor and FKBP12) in antigen-specific CD8 T 
      cells. In our protocol, a mouse model of lymphocytic choriomeningitis virus 
      infection is used, but the methods can be extended to other viral and bacterial 
      infections as well as vaccinations. Also, the similar approach can be applied to 
      analysis of CD4 T-cell responses.
FAU - Araki, Koichi
AU  - Araki K
AD  - Department of Microbiology and Immunology, Emory Vaccine Center, Emory University 
      School of Medicine, Atlanta, GA, USA. karaki@emory.edu
FAU - Konieczny, Bogumila T
AU  - Konieczny BT
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Adaptor Proteins, Signal Transducing)
RN  - 0 (Carrier Proteins)
RN  - 0 (Regulatory-Associated Protein of mTOR)
RN  - 0 (Rptor protein, mouse)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - EC 5.2.1.- (Tacrolimus Binding Protein 1A)
SB  - IM
MH  - Adaptor Proteins, Signal Transducing
MH  - Animals
MH  - CD8-Positive T-Lymphocytes/cytology/*immunology
MH  - Carrier Proteins/genetics/metabolism
MH  - Cell Differentiation
MH  - Cell Line
MH  - Disease Models, Animal
MH  - Flow Cytometry
MH  - Gene Expression Regulation
MH  - *Gene Knockdown Techniques
MH  - Gene Silencing/immunology
MH  - Genetic Vectors
MH  - Humans
MH  - Immunity, Innate/*genetics
MH  - Lymphocytic Choriomeningitis/genetics/metabolism/virology
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - *RNA Interference
MH  - Regulatory-Associated Protein of mTOR
MH  - Retroviridae
MH  - TOR Serine-Threonine Kinases/*genetics/*immunology
MH  - Tacrolimus Binding Protein 1A/genetics/metabolism
MH  - Transduction, Genetic
EDAT- 2011/11/30 06:00
MHDA- 2012/03/17 06:00
CRDT- 2011/11/30 06:00
PHST- 2011/11/30 06:00 [entrez]
PHST- 2011/11/30 06:00 [pubmed]
PHST- 2012/03/17 06:00 [medline]
AID - 10.1007/978-1-61779-430-8_19 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2012;821:305-16. doi: 10.1007/978-1-61779-430-8_19.