YFP shRNA

Design of a viral vector for knocking down Yellow Fluorescent Protein expression (and whatever it is attached to).

To rescue limited brain regions in the FKO mice, we will construct a rAAV virus which will express a short-hairpin RNA (shRNA) that causes degradation of YFP, using previously published shYFP sequences PMID 16676321 PMID 18708075. PMID 15582578 ( Targeted degradation of the 771-nt YFP portion of a YFP-tagged transgene should be sufficient to block expression of the complete transgene PMID 16676321.

The construct will also contain DsRed or mCherry as the reporter gene to allow co-localization with the constitutive YFP reporter in 14-3-3 FKO mice. A rAAV-DsRed vector without shYFP will be constructed as a control virus.

Oligonucleotides encoding the sense and antisense strands of shRNA against GFP/YFP PMID 15582578:

siGFP sense: 5’-gat ccc gca agc tga ccc tga agt tca ttt caa gag aat gaa ctt cag ggt cag ctt gct ttt ttg gaa a-3’

siGFP anti-sense: 5’-agc ttt tcc aaa aaa gca agc tga ccc tga agt tca ttc tct tga aat gaa ctt cag ggt cag ctt gcg a-3’