CSB349L16 - Length of 3 UTR as well as its polyA helping...

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Length of 3’ UTR as well as it’s polyA helping stabilizing it for translation. Position where transcription cleavage occurs as well as poly A tail. The sequence regions (AAUAAA), help recruit complexes for cleavage and polyA: CPSF, CstF and other co-factors which include PAP. PAP has two phases: slow and fast. b/c adding the As are slow until PABPII binds when there will be 200-250 A residues added. One of the few transcripts which aren’t PolyAed are the core histone transcripts. Variant histones have primary transcripts which are PolyAed though. Experimental techniques to look at this: Test tube w/ substrates and buffers. Determine the process over time . using combination of oligo-dt and RNase H. Regulating length of 3’ UTR can have a big impact on regulating protein function. E.g. BDNF genes. Alternative polyA can control the level of protein made!! A short UTR can still contain sequence to be polyAed or a longer UTR. You can shorten UTR to have it not have poly A (you’ll get small amount of protein made b/c these types of RNA as quickly degraded b/c they don’t have a polyA). Anti-sense inhibition: if you have a primary RNA is sense orientation, if you provide anti-sense copy of the mature RNA and hybridize it to its target, you’ll create a dsRNA and it won’t be able to make a protein.
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Does this work for cancer genes? The evidence is up in the air. It’s really hard to do. They tried it with c. elegans to replicate this. In a gene specific way, they provided the sense RNA ss and the antisense RNA causing the two to ds. The ds was the control for the experiment. But the control worked way better than the antisense strand. Very strange. Worms and in plants, the dsRNA gets processed into smaller bits of dsRNA leading to RNA degredation and protein translation inhibition.
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discovery of mechanism of RNA interference. Can dsRNA reduce levels of RNA? Knockdown? On LH side has 3 different mutant worms (unc 22, fem-1, and unc54). Unc= unco-ordinated or paralyzed worms. The mutant itself’s phenotype is on far right. Unc-22 are twitching worms.
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