PMID- 14576327
OWN - NLM
STAT- MEDLINE
DCOM- 20031209
LR  - 20190501
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 31
IP  - 21
DP  - 2003 Nov 1
TI  - Inducible shRNA expression for application in a prostate cancer mouse model.
PG  - e127
AB  - RNA interference (RNAi) is a powerful tool to induce loss-of-function phenotypes 
      by inhibiting gene expression post-transcriptionally. Synthetic short interfering 
      RNAs (siRNAs) as well as vector-based siRNA expression systems have been used 
      successfully to silence gene expression in a variety of biological systems. We 
      describe the development of an inducible siRNA expression system that is based on 
      the tetracycline repressor and eukaryotic RNA polymerase III promoters (U6 and 
      7SK). For proof of concept we selectively inhibited expression of two catalytic 
      subunits of the phosphatidylinositol 3-kinase (PI 3-kinase), p110alpha and 
      p110beta, by using vector-derived short hairpin RNAs (shRNAs). Stable pools of 
      human prostate cancer cells (PC-3) exhibiting reduced levels of both PI 3-kinase 
      catalytic subunits due to the expression of corresponding shRNAs in an inducible 
      fashion were established and analyzed for their invasive potential in vitro as 
      well as in an orthotopic metastatic mouse model. This inducible system for RNAi 
      allows an unbiased and comparable analysis of loss-of-function phenotypes by 
      comparing selected isogenic cell populations on the induced and non-induced 
      level. In addition, conditional RNAi allows the study of essential and 
      multifunctional genes involved in complex biological processes by preventing 
      inhibitory and compensatory effects caused by constitutive knockdown.
FAU - Czauderna, Frank
AU  - Czauderna F
AD  - Atugen AG, Otto Warburg Haus (No. 80), Robert-Roessle-Strasse 10, 13125 Berlin, 
      Germany.
FAU - Santel, Ansgar
AU  - Santel A
FAU - Hinz, Michael
AU  - Hinz M
FAU - Fechtner, Melanie
AU  - Fechtner M
FAU - Durieux, Birgit
AU  - Durieux B
FAU - Fisch, Gerald
AU  - Fisch G
FAU - Leenders, Frauke
AU  - Leenders F
FAU - Arnold, Wolfgang
AU  - Arnold W
FAU - Giese, Klaus
AU  - Giese K
FAU - Klippel, Anke
AU  - Klippel A
FAU - Kaufmann, Jorg
AU  - Kaufmann J
LA  - eng
SI  - GENBANK/X05490
SI  - GENBANK/X07425
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Protein Subunits)
RN  - 0 (RNA, Small Interfering)
RN  - EC 2.7.1.- (Phosphatidylinositol 3-Kinases)
RN  - EC 2.7.7.6 (RNA Polymerase III)
RN  - F8VB5M810T (Tetracycline)
SB  - IM
MH  - Animals
MH  - Catalytic Domain
MH  - Cell Line, Tumor
MH  - *Disease Models, Animal
MH  - *Gene Expression Regulation, Neoplastic/drug effects
MH  - Humans
MH  - Male
MH  - Mice
MH  - Molecular Sequence Data
MH  - *Nucleic Acid Conformation
MH  - Phosphatidylinositol 3-Kinases/biosynthesis/chemistry/genetics
MH  - Promoter Regions, Genetic/genetics
MH  - Prostatic Neoplasms/enzymology/*genetics
MH  - Protein Subunits/biosynthesis/genetics
MH  - RNA Interference
MH  - RNA Polymerase III/genetics
MH  - RNA, Small Interfering/chemistry/*genetics/*metabolism
MH  - Tetracycline/pharmacology
PMC - PMC275484
EDAT- 2003/10/25 05:00
MHDA- 2003/12/10 05:00
PMCR- 2003/11/01
CRDT- 2003/10/25 05:00
PHST- 2003/10/25 05:00 [pubmed]
PHST- 2003/12/10 05:00 [medline]
PHST- 2003/10/25 05:00 [entrez]
PHST- 2003/11/01 00:00 [pmc-release]
AID - gng127 [pii]
AID - 10.1093/nar/gng127 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 2003 Nov 1;31(21):e127. doi: 10.1093/nar/gng127.