|shRNA Shared Technology Resource|
David P. Turner PhD
Assistant Professor, Department of Pathology & Laboratory Medicine
Location: BEB 422
Short hairpin RNA’s (shRNAs) can efficiently suppress gene expression in human cells. The shRNA Shared Technology Resource will provide investigators at MUSC access to genome wide human and mouse libraries that together encode a total of almost 160,000 shRNA clones against over 41,000 genes. The resource utilizes The RNAi Consortium’s (TRC) genome-wide lentiviral mouse and human libraries and investigators will have the option of ordering shRNA’s targeting single or multiple genes, gene family sets as well as pathway specific pooled libraries. The library will allow access to multiple shRNAs for a single gene which is important for validation against off target effects. This technology holds tremendous power, and is ready to help investigators at MUSC work toward breakthrough discoveries. The resource is housed in the Bioengineering Building (BE422) within the James E. Clyburn Research Center on Jonathan Lucas Street.
shRNA Library Features
- Largest and most validated shRNA collection
- Human Library: 20,018 genes, 129,695 clones (1500-96 well plates)
- Mouse Library: 21,171 genes, 118,062 clones (1374-96 well plates)
- Hairpins comprised of a 21mer base stem and a 6 base loop designed against NCBI REFSEQ
- Sequence, specificity & position scoring with the Broad Institute algorithm
- A minimum of 3-5 shRNA constructs are created for each target gene to provide varying levels of knockdown and to target different regions of mRNA transcript
- For any given RefSeq, there is often a shRNA clone targeting the 3'UTR for use in phenotypic rescue studies using cDNA expression constructs.
Lentiviral Vector Features:
- shRNA cloned into the pLKO vector developed by the Broad Institute
- Allows for both stable or transient transfection
- self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids
- Stable gene silencing is selected using the puromycin selectable marker
- Integrates for long-term knockdown
- Transduces virtually any cell type (dividing or non-dividing)
- No interferon response
- Lack of recombination issues
Ordering shRNA’s of Interest
To inquire about or access the services offered by the shRNA Shared Technology Resource, please contact the director (contact information listed above).
Prior to services being performed, the requestor must submit an Order Form. Upon receipt, the requestor will be contacted, the request reviewed and if required additional information requested before pricing is provided. Please note: all requests must be supported by a current investigator IBC# covering the target genes of interest and the use of lentiviral particles.
If you publish a manuscript including research supported by the shRNA Technology Resource, the HCC recommends the following text to be placed in the acknowledgement section:
"Supported in part by the shRNA Shared Technology Resource, Hollings Cancer Center, Medical University of South Carolina."
►If you have any questions please e-mail Dr David P. Turner at email@example.com