PSF-U6 - SIRNA/RNAI EXPRESSION PLASMID

The expression of short hairpin RNA in eukaryotic cells. The U6 is transcribed by RNA polymerase III (RNA pol III) which terminates transcription at a string of thymidine residues typically a minimum of five residues in mammalian cells. Unlike most of our vectors, this vector contains an RNA pol III terminator downstream of the XhoI site although it is more common to insert a terminator sequence at the end of the sequence you insert into the MCS that encodes a shRNA.

Promoter Expression Level:

Multiple cloning site notes: This vector has a modified MCS compared to standard plasmids. The ATG start codon in the NcoI site has been removed as have the Shine-Dalgarno and KOZAK sequences as these are not required for RNA Pol3 expressed transcripts.

The terminator for RNA polymerase III has been positioned immediately downstream of the XhoI site. There is an additional string of Thymidine residues (the terminator sequence for RNA Pol III) at the end of the SV40 polyA region however using this site will produce a long transcript and has not been validated. We recommend inserting your hairpin DNA sequence between Hind3 and XhoI. The theoretical RNA position +1 is the first base of the NotI restriction site.

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