The Lentiviral Laboratory

About the Lab

The USC School of Pharmacy Lentiviral Laboratory, located on the second floor of the Pharmaceutical Sciences Center, 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.

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 lab 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.

High level of expression that can be achieved using the rLV method in neurons

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.

Functionality of P2X4 receptors as determined using whole-cell patch-clamp techniques

Image illustrations: Recombinant lentiviral (rLV)-mediated P2X4 Transduction of Rat Hippocampal Neurons (10X). The image illustrates the high level of expression that can be achieved using the rLV method in neurons. The Data image illustrates the functionality of P2X4 receptors as determined using whole-cell patch-clamp techniques. ATP (agonist) activated currents of P2X4Rs expressed in hippocampal neurons using lentiviral technology are inhibited by ethanol (10μM ATP-induced currents ± 50mM ethanol).

Resources

The Lentiviral Laboratory is a well equipped biosafety level 2+ lentiviral cell cuture room. This cell culture room contains a Class II type A2 culture hood for sterile work (Nuaire, Plymouth, MN), two cell culture incubators (Sheldon Manufacturing, Cornelius, OR), inverted light microscope with fluorescence capabilities (Carl Zeiss, Inc. Thornwood, NY), table top centrifuge (Eppendorf, Hamburg, Germany), refrigerator, water baths and other commonly used instruments.

Services provided include: 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.

Chargebacks

The Lentiviral Laboratory provides viruses on a fee-for-service basis and forms 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 $3,000 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 lab 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.

Services turnaround time:

  • 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

Scheduling

Dr. Asatryan coordinates the lab schedule. Her contact information is below.

Contact Us

Lab location:
PSC 210BB
Health Sciences Campus
USC School of Pharmacy
1985 Zonal Avenue
Los Angeles, CA 90033

Daryl L. Davies, PhD
Associate Professor of Clinical Pharmacy
Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics & Policy
Director, Alcohol and Brain Research Laboratories
USC School of Pharmacy, PSC500
(323) 442-1427
ddavies@usc.edu

Liana Asatryan, PhD
Senior Research Associate
Titus Family Department of Clinical Pharmacy and Pharmaceutical Economics & Policy
USC School of Pharmacy, PSC400
(323) 442-1554
asatryan@usc.edu

Letisha R. Wyatt
Graduate Student
Department of Pharmacology and Pharmaceutical Sciences
USC School of Pharmacy, PSC500
(323) 442-1426
lrwyatt@gmail.com