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The lentivial core facility provides services for production of small scale lentiviral stocks containing the gene of interest, lentiviral titrations, concentrations and transductions of cells of interest. We also construct individual lentiviral vectors including sub-cloning of the gene of interest into a pLVX-puro lentiviral vector.
RNA Interference (RNAi) is one of the most important technological breakthroughs in modern biology, allowing us to directly observe the effects of the loss of function of specific genes in mammalian systems. To this end, we are currently developing a variety of protocols for conducting loss of function studies in mammalian cells and animal models using RNAi technologies via the use of siRNA, shRNA or miRNA expression in conjunction with lentiviral delivery methods.
SERVICES OFFERED: 1) Providing viruses strictly on a fee-for-service basis and 2) Forming collaborations that lead to grant subcontracts/co-investigator status. Several examples of fees that may be encountered are provided only as examples. Much of the costs are with the understanding that this is a collaborative venture which requires input and feedback from the PI. If a USC School of Pharmacy laboratory hands us the cDNA in a plasmid, we will charge $3000 per new construct and $500 for each new batch of existing virus. If we need to start by PCR to obtain the cDNA, then we add about $750; more for long cDNAs. Any direct costs such as purchasing vectors with the cDNAs or obtaining oligos to generate miRNAs are charged. These calculations are based on amounts to help offset some of the personnel costs associated with the core facility but do not include equipment costs or the time that we spend in designing the vector, troubleshooting cloning, etc.—we usually consider these services as part of our collaboration with USC Pharmacy investigators. Charges for Investigators outside of School of Pharmacy are approximately 25% higher.
Lentiviral vector construction – 4-6 weeks
Lentivirus packaging and concentration (small scale) – 5 days
Lentiviral stock titrations – 2 weeks
Lentiviral infections of primary cells including neurons – 4-10 days
Our goal is to facilitate research studies by providing lentiviral stocks for delivery and expression of transgenes in difficult to transduce primary cells and/or tissue culture. High titers of lentivirus that are produced by the core provide high efficiency and long-term expression of transgenes. Very efficient in production of stable cell lines. The lentiviral delivery of siRNA or shRNA that we are currently developing has also proven very effective for gene silencing applications.
Core Services: Lentiviral vector construction. Backbone vectors for cDNA expression. Small-scale virus production for gene transfer into cell lines. High-titer virus production for gene transfer into primary cells and in vivo. Production of stable cell lines. Consultation for vector modalities, troubleshooting, support documentation for grants. Future services will address the need for RNA interference.
Contact us if this is something you are interested in embracing in your research program.
Titus Family Department of Clinical Pharmacy and
Pharmaceutical Economics & Policy
School of Pharmacy
University of Southern California
1985 Zonal Avenue, PSC 210BB
Los Angeles, CA 90033
Liana Asatryan, PhD
Senior Research Associate
Titus Family Department of Clinical Pharmacy and
Pharmaceutical Economics & Policy
School of Pharmacy
University of Southern California
1985 Zonal Avenue, PSC 400
Los Angeles, CA 90033
(323) 442-1554; FAX (323) 442-1704; e-mail asatryan@usc.edu
Letisha R. Wyatt
Graduate Student
Department of Pharmacology and Pharmaceutical Sciences
School of Pharmacy
University of Southern California
1985 Zonal Avenue, PSC 500
Los Angeles, CA 90033
(323)442-1426; FAX (323)442-1704; e-mail lrwyatt@gmail.com
Daryl L. Davies, Ph.D.
Associate Professor of Clinical Pharmacy
Titus Family Department of Clinical Pharmacy and
Pharmaceutical Economics & Policy
Director, Alcohol and Brain Research Laboratories
School of Pharmacy
University of Southern California
1985 Zonal Avenue, PSC 500
Los Angeles, CA 90033
(323) 442-1427; FAX (323) 442-1704; e-mail ddavies@usc.edu
You can contact:
Liana Asatryan, PhD
asatryan@usc.edu
Tel:(323)442-1554