This Request for Proposal (“RFP”) is an invitation to prospective Proponents to submit Proposals for the provision of Open Reading Frame (ORF) libraries, in gateway expression construct format, as they offer an unprecedented way to interrogate the gain-of-function of virtually any gene in a highly flexible manner. The University’s researchers intend to establish a streamlined pipeline for the generation of ORF constructs that are amenable for multiple purposes, including: 1) high titer viral expression in difficult to transfect primary mammalian cells; 2) bacterial expression for protein purification; and 3) fluorescent tagging for biochemical and cell biological manipulation in culture. This pipeline will ultimately be integrated in the University’s forthcoming gene-manipulation core to offer this service to the broader scientific community.
General Description:
ORF libraries targeting the majority of genes in the human genome in expression ready format for multiple outputs, including but not limited to: 1) viral vector expression; 2) bacterial expression via GST- or His6-expression; and 3) mammalian expression with GFP fusion tag. The library must be in application-ready viral and plasmid stock or contain a production package of sufficient size for the end-user to prepare the viral library. ORF constructs must be sequence verified and amenable to both N- and C-terminal tagging.
Mandatory Minimum Requirements:
1. Genes Targeted
1.1ORFeome must target the majority (>10,000) of expressed genes.
1.2Must express human genome.
1.3Collection of genes must be sequence-verified.
1.4ORFs must be amenable to N- and C-terminal tagging.
2.Expression Ready
2.1Must include a glycerol stock back-up in 96-well plate format.
2.2Must include application-ready viral and plasmid stock or a production package of sufficient size for end-user to prepare 10 µg of plasmid and 10 mL of virus for each vector.
3. Other Requirements
3.1Shipping on dry ice must be included and the supplier must cover risk of damage and loss during shipment transit time up to the University’s Receiving location.
3.2Must include one (1) year warranty on bacterial colony viability.
