Conditional gene knock-down by CRE-dependent short interfering RNAs.
Short interfering RNAs (siRNAs) are short (21-23 nt) double-stranded RNAs that direct the sequence-specific degradation of corresponding mRNAs, resulting in suppression of gene activity. siRNAs are powerful tools for gene functional analysis in mammals. Chemically synthesized siRNAs permit transient gene repression but preclude inhibition of stable gene products as well as long-term phenotypic analyses. Permanent gene suppression can be achieved by transcribing siRNAs as stem-loop precursors from Pol III promoters. This approach, however, has a major limitation: inhibition cannot be controlled in a time- or tissue-specific manner. Thus, the approach cannot be applied to genes essential for cell survival or cell proliferation. To overcome these limitations, we have designed a CRE-lox-based strategy that allows one to repress gene activity in a time-dependent manner in cells, and in a time- or tissue-dependent manner in animals. Our approach promises to improve dramatically the procedures for functional genetics in mammals.
Fritsch
Lauriane
L
UPR 9079 CNRS-Ligue Nationale Contre le Cancer, UPR 1983 CNRS, Institut André Lwoff, Villejuif, France.
Martinez
Luis A
LA
Sekhri
Redha
R
Naguibneva
Irina
I
Gérard
Mathieu
M
Vandromme
Marie
M
Schaeffer
Laurent
L
Harel-Bellan
Annick
A
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004258:DNA Polymerase III / Q000235:genetics* / Q000378:metabolism*; D016158:Genes, p53 / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D009132:Muscles / Q000166:cytology / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D034741:RNA, Small Interfering; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases; D004258:DNA Polymerase III