pVALIUM22 allows cloning of short hairpins as annealed synthetic oligos, targeted against specific genes via the production of short hairpin microRNAs; contains v+t1.8 as a selectable marker; the P-transposase core promoter; a multiple cloning site (MCS) for cloning shRNAs in a mir-1 scaffold; a ftz 3'UTR intron followed by a fs(1)K10 3'UTR to facilitate shRNA processing and export from the nucleus; two pentamers of UAS, one of which is flanked by loxP sites and can thus be excised using the P1cre recombinase to generate a 5xUAS derivative; two gypsy insulator sequences flanking the transcription unit to enhance hairpin DNA transcription; an attB sequence to allow phiC31:int-targeted integration at genomic attP landing sites.
Efficacy of pVALIUM20-based vs pVALIUM22-based RNAi constructs in germline and soma compared. pVALIUM20 generates effective knockdown phenotypes in both germline and soma; pVALIUM22 is more effective for knockdowns in the germline and effects appear to be restricted to the female germline (excluding somatic ovarian cells), but is significantly less effective in the soma.
See TRiP(http://fgr.hms.harvard.edu/trip-plasmid-vector-sets) for downloadable maps, sequences and cloning protocols.
'VALIUM' stands for Vermilion-AttB-Loxp-Intron-UAS-MCS. pVALIUM22 triggers effective RNAi knockdown and corresponding phenotypes in the female germline; it is less effective in somatic cells.