PMID- 22125078
OWN - NLM
STAT- MEDLINE
DCOM- 20120316
LR  - 20211203
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 821
DP  - 2012
TI  - Assessing the function of mTOR in human embryonic stem cells.
PG  - 361-72
LID - 10.1007/978-1-61779-430-8_23 [doi]
AB  - We described a protocol for dissecting the function of an important 
      serine/threonine protein kinase, mammalian target of rapamycin (mTOR), in 
      regulating the long-term undifferentiated growth of human embryonic stem cells 
      (hESCs). The function of mTOR in hESCs was inactivated with a highly specific 
      chemical inhibitor, rapamycin, and gene-specific small-hairpin RNAs, and the 
      effects were evaluated under self-renewal or early differentiation conditions. We 
      found that inactivation of mTOR impairs proliferation and enhances mesoderm and 
      endoderm activities of hESCs. This protocol described a general strategy for 
      studying the function of key genes and signaling events during hESC long-term 
      self-renewal and early lineage specifications with pharmacological and genetic 
      approaches.
FAU - Zhou, Jiaxi
AU  - Zhou J
AD  - Department of Cell and Developmental Biology, Institute for Genomic Biology, 
      University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA. 
      jiaxi@life.uiuc.edu
FAU - Li, Dong
AU  - Li D
FAU - Wang, Fei
AU  - Wang F
LA  - eng
GR  - GM083601/GM/NIGMS NIH HHS/United States
GR  - GM083812/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (RNA, Small Interfering)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - W36ZG6FT64 (Sirolimus)
SB  - IM
MH  - Animals
MH  - *Cell Differentiation
MH  - Cell Lineage/drug effects
MH  - Cell Proliferation/drug effects
MH  - Embryonic Stem Cells/*cytology/*enzymology
MH  - Endoderm/cytology/drug effects
MH  - Gene Knockdown Techniques
MH  - HEK293 Cells
MH  - Humans
MH  - Mesoderm/cytology/drug effects
MH  - Mice
MH  - RNA, Small Interfering/genetics
MH  - Signal Transduction
MH  - Sirolimus/pharmacology
MH  - TOR Serine-Threonine Kinases/*antagonists & inhibitors/genetics/*physiology
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_23 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2012;821:361-72. doi: 10.1007/978-1-61779-430-8_23.