Seven pSico plasmids are available from Addgene:
- pSico, pSicoR, pSico p53, pSico p53R (published)
- pSico PGK puro, pSicoR PGK puro, pSico PGK EGF (unpublished)
Find Jacks Lab Plasmids
Maps & Sequences
- pSico Map
- pSicoR-GFP Map
- pSico Genbank Sequence
- pSicoR-GFP Genbank Sequence
- pSicoR-pgk Puro Genbank Sequence
pSico Oligo Design
The Ventura Lab at MSKCC has pSicoOligomake 3.0. Please see their website for further information.
Use the Mac OSX program pSicoOligomaker 1.5 to select and design oligos for pSico and plentilox3.7.
To operate it, simply paste your sequence in the "sequence" window, give it a name (optional), select a cutoff value (from -2 to 9) and click "search". The program will list in the right window all the potential 19-mers with a score equal or higher than the cutoff. The score is given based on a set of criteria published in Nature Biotech. by Reynolds et al. in 2004. According to the authors, a 19-mer with a value higher than 6 has approximately 90% chance of achieving silencing of the target mRNA.
You can select all targets and paste them in Word or Excel to store the result. In addition, if you type or paste a 19-mer in the field "target" and click on "make oligos" the program returns the sequence of the forward and reverse oligos (both in the 5' to 3' orientation) that have to be ordered to clone in pSico. The button "order" directs you to the web page we use to order oligos. The button "settings" lets you change the criteria to design the oligos (loop sequence and length, overhangs, etc) so that it can be used to design oligos for other vectors.
"Clear sequence" tells the program to remove all illicit characters, including spaces and "end of line", from the sequence. In addition, it converts the sequence to all caps.
In case you don't have a Mac, or you prefer to design your oligos manually, here is an example:
How to clone into pSico and pSicoR
We are using the lentiviral packaging system made by Inder Verma's Lab (at the Salk). We have an MTA for his vectors, so we are not allowed to send those out to others. You can look on his lab's web site or papers to find the vector information; he may have more updated versions than we have now. You can also look for the information in the reference cited in the Ventura paper (Rubinson, D. A., Dillon, C. P., Kwiatkowski, A. V., Sievers, C., Yang, L., Kopinja, J., Rooney, D. L., Ihrig, M. M., McManus, M. T., Gertler, F. B., et al. (2003) Nat. Genet. 33, 401 406.) which made the lentilox 3.7 that pSICO is based on.
RNAi Resources (pLL3.7 Information Pages)
- Cloning of shRNAs into pLL3.7 vector
- Production of lentivirus
- Lentiviral RNAi Vectors
- Lentiviral Transgenics:
- Lentiviral Chimeras