Viral Vector Core (VVC)

 

How to acknowledge us in your publications? University of Maryland School of Medicine’s Viral Vector Core – Baltimore, Maryland.

Overview of Services

MISSION

UMB Virus Vector Core (VVC) offers Adeno Associated Virus (AAV) and lentivirus (LV)-based gene delivery systems to be used in gene therapy for targeting a gene of interest (GOI) in in-intro and in-vivo studies. The VVC offers services for custom gene cloning and packaging in AAV or LV. The mission of this Core is to provide high-quality viral vectors, reducing the time and cost for the investigators utilizing modern tools and technology. The Core offers free consultancy for gene construct designing strategy, discussing the research project, implementation, and troubleshooting.

SERVICES OFFERED

  1. The Viral Vector Core generates ready-to-use pre-made viruses that are available for immediate pick-up.
  2. Production of custom vectors – “from scratch” designed according to the needs and specifications of the investigator.
  3. A wide selection of AAV serotype and purification grades for in-vivo (live tissue) or in-vitro (primary cell lines).
  4. CRISPR-based genome editing, vector cloning, guide RNA designing, and screening in cell lines. Cloning of CRISPR effector complexes (CRISPRi and CRISPRa) and packaging into AAV or Lentivirus.
  5. The Core will offer consultation service on the design of viral vectors, providing help in choosing virus type and serotype optimal for the cell- and tissue-specific region.

AAV virus

AAV virus is packaged by using triple transfection in HEK393T cells. Helper plasmid, red/cap plasmid, and transgene plasmid. After 72hr of transfection, the virus is purified by ultracentrifugation method. 

Titer- We are routinely providing a very high titer range from 1012  to 1013 GC/ml, Note- titer may depend on the loading size of the target gene and serotype.

Quality control

  1. DNA sequencingof target gene
  2. Titer- Titer of the virus tested by qPCR using standard. 
  3. Purity-AAV samples are routinely inspected by SDS PAGE followed by color staining. This assay confirms to see the AAV capsid protein VP1, VP2, and VP3 molecular weight and relative density. Note-We don’t test this QC for all the viruses we made but we do whenever we needed.
  4. Endotoxin Free-To avoids the endotoxin-free AAV or LV virus. We used an Endotoxin-free DNA extraction Kit (Qiagen).

Lentivirus

UMB Viral Vector Core generates the lentivirus using a second-generation lentivirus system. We are using standard psPAX2 and envelop protein CMV-SVS-G/ pMD.2G plasmid transfected into HEK293 cell line. Lentivirus containing clear supernatant concentrated by ultracentrifugation. Our service provides in-vivo and in-vitro grade ultra-purified lentivirus.

Lenti titer- We used Lenti-X™ qRT-PCR Titration Kit (TAKARA) or used a standard method to determine the LV concentration.

 

Service                                                                                            Amount/Titer                Internal Price      External Price    Corporate price

Lentivirus

Lentiviral purified supernatant                                                        10-20ml, 107 – 108 TU/ml          $500.00            $900.00

Ultra-purified in-vivo grade concentrated Lentivirus                          1000µl, 108–10TU/ml               $1450.00          $1550.00              $1950

 

AAV virus

AAV Customize virus packaging                                                      700-1000ul, ~1013GC/ml       $2050.00         $2450.00               $3150

AAV stock virus                                                                            100µl, ~1013 GC/ml                $320.00           $350.00                 $380.0

  NOTE-

1-All 'internal' and 'external' users please sign in iLab system to pay the bill. 

2-All the above cost subjects are to update every financial year on July 1.

3-Please deposite/ship the samples at 655 West Baltimore Street, BRB Rm7018, Baltimore, 21201

Leadership

Ramesh Chandra Director 410 706 2770 rchandra@som.umaryland.edu

 Website

https://www.medschool.umaryland.edu/cibr/core/viralvector

 

Support the Cores that Support Your Research.  

 

Contacts

Name Role Phone Email Location
Ramesh Chandra, PhD
Director
 
410 706 2770
 
RChandra@som.umaryland.edu
 
655 West Baltimore Street, BRB Rm7018, Baltimore,21201
 

Service list


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