The Roswell Park Cancer Institute/University at Buffalo shRNA Core shared resource serves as the focal point of RNAi expertise for the RPCI/UB research communities. The shRNA resource provides a centralized service that supports investigators interested in gene transfer using viral vector technologies.
REAGENTS AND SERVICES AVAILABLE
shRNA libraries purchased from Thermo Scientific Open Biosystems:
- Expression Arrest pGIPZ Lentiviral shRNAmir-GFP Human shRNA Library and the Expression Arrest pSM2 Retroviral shRNAmir Human and Mouse shRNA Libraries
~62K unique shRNAmir constructs targeting the entire genomes (>25,000 genes) with ~3-5 constructs/target
- Expression Arrest pSM2 Retroviral shRNAmir Human Cancer shRNA Library
2311 shRNAmir constructs targeting 967 human cancer related genes
- Decode™ RNAi pGIPZ Lentiviral Screening Pools
The high titer viral pool format enables genome-scale RNAi screening without the cost and labor associated with individually arrayed screens.
The shRNA libraries purchased from Open Biosystems contain microRNA-adapted shRNA (shRNAmir) constructs that are modeled after primary microRNA transcripts allowing efficient processing via the endogenous microRNA pathway and producing increased, more specific knockdown. Designed with features aimed at increasing gene knockdown efficiencies and versatility of use in RNAi screens, shRNA libraries provide solutions for diverse RNAi applications. In addition, the libraries have an incorporated DNA “bar-coding” system, by which each RNA molecule has been tagged with a unique 60 nucleotide DNA sequence. This facilitates shRNA validation and gene identification after library screens.
shRNA libraries purchased from Cellecta:
- DECIPHER™ Pooled shRNA Human and Mouse Lentiviral Libraries from CELLECTA
These Bar-coded lentiviral shRNA libraries are optimized for RNAi Genetic Screens in a pooled format. They were made to cover the entire annotated human and mouse gene sets with a high percentage of functionally validated sequences and inclusion of a redundant set of 5 to 6 shRNA per gene. The DECIPHER libraries are broken down into modules: three modules for human, and two modules for mice. Each module targets roughly 5,000 genes--variously associated with diseases, known drug targets, or signaling pathways--and represents over 25,000 unique shRNAs. This modular format was designed to help eliminate experimental concerns associated with large-scale screens and provides a more comprehensive approach to screening in general. In addition, the libraries use optimized 18-nt bar-codes that facilitate the identification of functional shRNAs and high-throughput sequencing data analysis using the Illumina platform.
Mycoplasma testing is available in the shRNA Core and uses the Agilent Mycoplasma Plus PCR Primer Set.
Irwin Gelman, PhD
Voice: (716) 845-7681
Using the Resource
For consultation and questions reagrding experimental design, please contact Dr. Gelman. Investigators interested in utilizing the services of the shRNA Resource should contact Renae Holtz.
Voice: (716) 845-1585
The shRNA Resource is located in the Center for Genetics and Pharmacology (CGP), Room L2-135. Hours of operation are weekdays, 8:30 am - 4:30 pm.