The shRNA and ORF reagents are available to The University of Texas MD Anderson Cancer Center researchers ONLY due to licensing agreements. If you have any questions, please contact Yutong Sun, Ph.D. at firstname.lastname@example.org.
Individual clone distribution as glycerol stocks and plasmids
FGC distributes individual shRNA or ORF in forms of glycerol stock (in LB medium containing 8% glycerol) or plasmid (midi-prep, >100ug). ORF plasmids will be verified by partial sequencing before they are released to researchers. The turnaround time is 2-4 business days for the glycerol stock and 3-5 business days for the plasmids.
Individual Lenti- or retro-virus packaging and transduction
FGC conducts plasmid purification, lenti- or retro-virus packaging and titer measurements. We also conduct this service if researchers provide us with their own viral vectors. The turnaround time is 4-6 weeks.
Customized Sub-libraries (glycerol stock, plasmids and/or lentivirus)
Based on the needs of researchers, FGC will “cherry-pick” shRNA or ORF glycerol stocks in a 96-well plate format. Depending on the researchers’ requests, we will purified the plasmids (transfection compatible) in 96-well plate format, and/or conduct lentiviral packaging in 96-well plates.
Establishing Somatic Gene-knockout Cell lines using the CRISPR/Cas9 technology
FGC will provide multiple clones of somatic gene-knockout cells and will validate gene-knockout by PCR (HR method) or sequencing (NHEJ method). Requirements: 1) cell line can tolerate clonal selections; 2) Loss of target gene will not kill the cells or significantly decrease cell proliferation rate; 3) cell line is compatible with transfection or transduction; 4)Western blots (for proteins), qPCR (for non-coding RNAs) or other assays are available to detect the loss of a target gene.
FGC staff will spend some of their efforts to facilitate MD Anderson Cancer Center researchers optimizing screen assays, perform screens and data analysis.
Custom shRNA or ORF sub-cloning
FGC subclones customized shRNAs into the pLKO.1 vectors, pGIPZ or pTRIPZ vector, or an ORF into an expression vector.
The following instruments are open to MD Anderson researchers: robots (Tecan EVO100 and EVO200) and High-content imaging systems (ImageXpress micro and ultra).
Pre-assembled shRNA libraries
FGC distributes the following pre-assembled shRNA libraries targeting the following gene families: Human kinases; Human phosphatases; and human E3 ligases. In addition, we also distribute Cellecta human Genome-Wide Pooled shRNA library (lentivirus).
TKO v3 CRISPR Library
FGC distributes the TKO v3 CRISPR Library (lentivirus, Dr. Moffat’s lab at the University of Toronto, https://www.addgene.org/pooled-library/moffat-crispr-knockout-tkov3/).