The goal of the Viral Vector Core Facility is to assist Institute investigators, as well as outside laboratories, with cost-effective production of high-quality, high-titer lentiviral, retroviral and adenoviral vector. The Viral Vector Core offers a variety of services including ready-to-transduce lentiviral and adenoviral particles, and custom lentiviral, retroviral, and adenoviral production and titrations. Packaging, concentrating, and titration of user-supplied lentiviral constructs are the major activities of the core. Our most recent “The Works” service includes custom lentiviral vector construction for constitutive or inducible cDNA/shRNA expression as well as promoter/enhancer reporter, viral packaging, and titration (ready-to-transduce control particles provided). Custom adenoviral productions including virus recovery, amplification, purification, and titration are now available. The Core is fully equipped for production of consistent quality viral vectors in carefully monitored producer cell lines (293T and 293A). The Core is also collaborating with the SBP Functional Genomics core facility to provide custom viral RNAi (shRNA) design and screening, and the animal core facility to provide custom viral transduction (in vivo). Consultation is provided on how to use the vectors and to address any other questions.
The core facility focuses on six major types of services:
1. Ready-to-transduce lentiviral preps – include: (1) constitutively expressed or inducible fluorescent protein, antibiotic selection marker, or bioluminescence reporter (2) nuclear or membrane localization tool (3) constitutively expressed non-silencing shRNA control.
- CMV-NLS-eGFP (nuclear eGFP)
- CMV-NLS-mCherry (nuclear mCherry)
- CMV-eGFP-CAAX (membrane eGFP)
- CMV-mCherry-CAAX (membrane mCherry)
- CMV-mCherry-CAAX-2A-NLS-eGFP (membrane mCherry & nuclear eGFP)
- LKO.1-scrambled shRNA (Sigma-Aldrich)
- GIPZ-scrambled shRNA (Open Biosystems)
- TRIPZ-scrambled shRNA (Open Biosystems)
2. Custom packaged lentiviral (HIV-1 and FIV) and retroviral production – is based on constructs provide by the customer. Titration is offered for virus ubiquitously expressing fluorescent marker.
3. Custom production of adenoviral particles (Ad5) from user-supplied construct or virus - includes: (1) adenovirus recovery (2) high purity adenoviral preparation using CsCl gradient purification (3) titration using the plaque assay.
4. "The Works" custom lentiviral vector construction – offers vector generation based on cDNA or shRNA of interests with a choice of constitutive/inducible expression vectors, viral production, and titration.
5. "The Works" custom lentiviral-adenoviral vector swapping – includes: (1) constitutive expression of custom cDNAs (2) inducible shRNA of interests expressed under a new Tet inducible system (3) Custom promoters/enhancer reporter vectors. When compared to lentiviral vectors, adenoviral vectors have several advantages such as (1) larger insert size (8-10 kB), (2) high titers (above 10E10 PFU/mL), (3) episomal (transient) expression.
6. Other Core services
- Custom viral particles concentration and purification with user-provided viral supernatants allows customers to increase viral titer (up to 1000 fold) based on crude viral supernatants.
- 100x viral concentration kit provides the concentration of crude viral lysates from small-scale viral production. Our kit is quick, easy, and allows for the long term storage of your virus at -80ºC.
- Large scale DNA plasmid preparation (ready for viral production) starts from a small amount of customer's vector (e.g. miniprep) or transformed bacterial stock.
- Ready-to-use zbFGF (zebrafish basic FGF) is a substitute for human bFGF on hES cell culture. High quantity and quality with less cost. Quality control is done batch by batch before releasing and the dilution of zbFGF is tested on hES cells.
- Ready-to-use polybrene - (1 mg/mL, 100µL per aliquot, 5 aliquots)
- Consultation is free for core users.
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.
For a Price List, please call (858)646-3100 ext. 4353 or email us.
Chun-Teng Huang earned his M.S. degree in microbiology at San Diego State University. With four years of research experience in virology, he was recruited by Sanford Burnham Prebys Medical Discovery Institute as manager of viral vector core facility in 2008. After establishing lenti-, retro-, and adeno-vector services, the core is now focusing on developing services related to non-segmented negative strand RNA virus.
Please call (858)646-3100 ext. 4353 or use the button below to send us an email.