The viral vector core facility at Sanford Burnham Prebys develops state-of-the-art viral vector-based gene delivery technology. Its portfolio ranges from lentivirus, retrovirus, adenovirus, AAV, VSV, sindbis virus, and Zika virus products as well as customized “The Works” viral vector construction and swapping service package. There are a variety of ready-to- transduce viral vector collections including constitutively expressed or inducible fluorescent proteins for nuclear or membrane localization and bioluminescence reporters for 3D drug screening. The viral core has integrated CRISPRs/Cas technology in the plasmid-free viral vector platform, providing scientists with choices of combined technologies to manipulate genes using CRISPR gene disruption, editing, inhibition, and activation. In addition, we extend technical expertise in nanoparticle research including exosome purification, analysis, and engineering. More importantly, the core actively participates and supports translational research to fight human diseases.
The Viral vector core facility focuses on the following services:
- Custom large scale rAAV production and titration. Serotypes include 1-9, 3B, 7M8, shH10, DJ, DJ/8, PHP.B, PHP.eB, PHP.S, B1, rh10, Anc80, and Retrograde. Ready-to-transduce rAAV-CMV-eGFP and CAG-NLS-eGFP/mCherry viruses are available upon request.
- Custom large scale lentiviral [HIV-1, integrase defective lenti-virus (IDLV), and FIV] and retroviral (MSCV and MMLV) production and titration is based on vector plasmid DNA provide by the user. Titration is offered for virus ubiquitously expressing fluorescence or puromycin selection marker.
- Ready-to-transduce lentiviral preps include: (1) doxy-inducible or constitutively expressed cytoplasmic fluorescent reporters w/out mammalian selection markers (2) nuclear or membrane localized fluorescent reporters (3) secreted or cytoplasmic bioluminescent reporters (4) doxy-inducible or constitutively expressed non-silencing control (scramble) shRNA (5) cell cycle and autophagy reporters.
- Custom large scale adenoviral particles (Ad5) production and titration from user-supplied construct or virus include: (1) adenovirus recovery (2) high purity adenoviral preparation using CsCl gradient purification (3) titration via plaque assay. Ready-to-transduce pAd5-eGFP, NLS/CAAX fused eGFP/mCherry, firefly or secreted gaussian/cypridina luciferase, and Cre recombinase viruses are available upon request.
- "The Works" custom lentiviral-adenoviral-rAAV vector construction and swapping include:
- doxy-inducible or constitutively expressed cDNA or shRNA
- promoter / enhancer reporter (3) microRNA sponges.
- siRNA to lenti shRNA vector conversion with choice of doxy-inducible H1 or constitutive hU6 promoter as well as puromycin or neomycin selection marker. Ready-to-transduce non-targeted siRNA converted or universal scramble shRNA lentiviruses are available upon request.
- Custom packaged Sindbis virus production and titration. Ready-to-transduce mCherryCAAX-eGFP virus, pSINREP5-based vector construction, packaging, and flow cytometry-based titration are available upon request.
- Custom VSV vector construction, production, and titration. Pseudotyped VSV-ΔG-eGFP with Ebola virus glycoprotein (EboV-GP) Zaire strain (Kissidougou-C15, 2014) or wild-type VSV glycoprotein (VSV-G) are available upon request.
- Custom Zika virus production and titration are available for African, Asian, and Brazilian (2015-2016 outbreak) strains.
Equipment & Resources
The Core is fully equipped with all the necessary equipment for production of consistent quality viral vectors including state-of-the art cell culture hoods and incubators under appropriate biosafety conditions, as well as an ultracentrifuge, microscopes and a dedicated -80°C freezer for storage of vectors. Producer cell lines 293T and 293A cells are routinely tested for the presence of mycoplasma using Lonza detection kit and discontinued at passage 20.
The Institute is in a partnership program with Sigma-Aldrich and Open Biosystems. Both companies provide affordable shRNA constructs. The Core also currently collaborates with Dr. Terskikh and Dr. Mercola labs to increase ready-to-transduce virus inventory.
Our lab uses the Cytocentric Xvivo System from BioSpherix Ltd to aseptically control O2, CO2, and temperature levels during the incubation, handling, and analysis of live cells without the disturbances which routinely occur in traditional cell culture incubators. This platform allows our core users to more accurately recapitulate physiologic, in vivo-like conditions during in vitro experiments with living cells. More information can be found at: biospherix.com/xvivo-system-model-x3
For a Price List, please call (858) 646-3100 ext. 4353 or email us.