Viral Vector Core
To provide preclinical grade, low cost, ready-to-use viral vectors to the CSMC research community for the advancement of basic and translational research efforts.
- Provide vectors of Lenti and Adeno viral origins.
- Evaluate and optimize vector constructs.
- Small or large scale vector production services including, packaging the vector constructs, concentrating viral particles, measuring viral vector titer (p24/ml, TU/ml) and quality control.
- Make available pre-made vectors carrying reporter genes, including fluorescent markers or luciferase.
- Under special circumstances, provide whole genome or pathway-specific cDNA or shRNA containing lenti vector libraries of human and mouse origins for high-content screening.
- ustom vector services including, cloning gene (cDNA) or shRNA of investigator’s interest into appropriate vector, sequence analysis, protein expression verification and vector production.
Director: Vaithi Arumugaswami, MVSc., PhD.
Email: Arumugaswami@cshs.org
Phone: 310 248 8584
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Panels show human cells (HEK-293) used for producing an adenovirus vector carrying a green fluorescent protein gene. A. Bright field microscopic view of infected cells showing virus-mediated cellular changes (Note: rounded up cells). B. Fluorescent microscopic view of panel A displaying fluorescent protein produced by adenoviral vector.
New User Information
Cedars-Sinai Medical Center Viral Vector Core (VVC) was established in 2011 to serve the viral vector production needs of CSMC researchers. We are located in the third floor of Steven Spielberg building SSB314 at Regenerative Medicine Institute. Currently, standard and custom vector production services are available for both lenti and first generation adenoviral vectors.
Info for ordering vector core services and prices can be found in the following sections.
The new users are highly encouraged to discuss with the VVC Director Dr. Arumugaswami regarding vector requirements. The investigators planning on using viral vectors and recombinant DNA should obtain the approval of Institutional Biosafety Committee (IBC) and other regulatory agencies prior to commencement of experiments.
Scheduling
Scheduling pick-up and delivery of plasmids, viral vectors and other reagents can be done by email or phone.
Vector Catalogs
1. Adeno Shuttle Vector 2. Lenti Vectors VVC
Ordering Services
For ordering, please download the VVC ORDER form and submit by email or fax.
Prices
Lenti Vector Rates:
| Service | Price | |
| Standard service: Virus packaging and concentrating with titration (Researcher provides vector) | 4 plates (15 cm dishes) | Contact: Dr. Xiong for pricing. |
| 8 plates (15 cm dishes) | ||
| *Vector plasmid construction only (one step cloning of gene or shRNA, screening, sequencing, plasmid preparation): |
*Price is for one step direct cloning of <2 kb insert; price for larger insert or multi-step cloning, please enquire.
Additional notes: 2-3 weeks turnaround time for vector production (for one step cloning custom vector production 3-4 weeks turnaround time). 4 plate scale titer is 1e6 - 1e7 in 200 ul.
Maximum insert size for lenti vector is <4 kb (including promoter, gene, polyA). Sub-optimum (larger size/toxic) inserts will adversely affect the yield.
Titer will be based on p24 ELISA.
Users will NOT be billed until virus is made. BSL2 biosafety containment required. Users should have biosafety approval through Institutional Biosafety Committee (IBC).
Adeno Vector Rates (1st gen):
| Yield pfu/ml (size) | Price | |
| *Ad plasmid construction, Concentrated AV packaging, with qualitative titration | 1e10 (2-3 ml) | Contact: Dr. Xiong for pricing. |
| Virus amplification | 1e10 (2-3 ml) |
*Quoted price includes one step direct cloning of <2 kb insert into shuttle vector; Higher price will apply for larger insert or multi-step cloning, please enquire for details.
Additional Notes: Maximum insert size is 7.5 kb.
Turnaround time- Vector DNA transfection and Viral amplification 5-8 weeks;
Custom vector cloning, DNA transfection and viral amplification WITH gene provided 8-12 weeks, and WITHOUT gene provided 12-16 weeks.
Users will NOT be billed until virus is made. BSL2 biosafety containment required. Institutional Biosafety Committee (IBC).
Protocols
Lenti vector:
For standard lenti production, we require a total of 100 microgram vector plasmid DNA at a concentration of 0.5 microgram per microliter. We recommend plasmid purification done by anion-exchange-column or cesium chloride gradients. The 260/280 nm absorbance ratio of 1.8 is required for optimum viral vector production. The researcher also has the option of providing bacterial stocks of the vector plasmid, and plasmid purification will be done by VVC staff (a separate fee may apply).
For custom lenti production, the researcher provides the gene or DNA of interest that needs to be cloned into a lenti vector plasmid. Please discuss with VVC staff.
First generation Adeno vector:
For amplifying adeno vector, the investigator provides a seed viral stock at a titer ranging 1e6 to 1e12 in a 10-100 microliter volume.
For custom adeno production, the researcher provides the gene template. VVC will clone the template into appropriate shuttle vector system. Please discuss with VVC staff.
Staff
Weidong Xiong, MD, MSc
Project Scientist,
Cedars-Sinai Medical Center
Contact Us
Mailing address:
8700 Beverly Blvd., SSB 319
CSMC Viral Vector Core
Los Angeles, CA 90048, USA
Email: weidong.xiong@cshs.org
Office Ph: (310) 967-0625
Lab Ph: (310) 248-8691
Fax: 310 248 8066
Links
National Gene Vector Biorepository (NGVB)
https://www.ngvbcc.org/AboutTheNGVB.action
Stanford University: Working with Viral Vectors
http://www.stanford.edu/dept/EHS/prod/researchlab/bio/docs/Working_with_Viral_Vectors.pdf
Addgene Lentiviral Protocols & Resources
http://www.addgene.org/lentiviral/protocols-resources/
News
2012
http://inewsletter.csmc.edu/CedarsScience/2012/May-17/Enlisting-viruses-for-a-good-cause.aspx
FAQ’s
What is the turn-around time for vector production?
- For lenti vector the turnaround time is 2-3 weeks (for one step cloning custom vector production 3-4 weeks turnaround time).
- For adeno vector: Standard viral amplification time is 5-8 weeks; Custom adeno vector cloning, DNA transfection and viral amplification WITH gene provided 8-12 weeks, and WITHOUT gene provided 12-16 weeks.
What is the typical vector yield?
- For 8 plate lenti vector, the anticipated yield is 1e6 - 1e8 per milliliter. The yield may be influenced by plasmid quality and nature of inserts (larger size/toxic).
- For the first generation adeno vector, the yield is ranging from 1e9 to 1e12 per milliliter.
What is the acceptable insert size?
- Maximum insert size for lenti vector is <4 kb (including promoter, gene, polyA).
- Maximum insert size for first gen adeno vector is 7.5 kb (including promoter, gene, polyA).
Open Positions
Currently no open positions.
