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

The Viral Vector Core generates ready-to-use pre-made viruses that are available for immediate pick-up.
Production of custom vectors – “from scratch” designed according to the needs and specifications of the investigator.
A wide selection of AAV serotype and purification grades for in-vivo (live tissue) or in-vitro (primary cell lines).
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.
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

DNA sequencingof target gene
Titer- Titer of the virus tested by qPCR using standard.
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.
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, 10⁸–10⁹TU/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

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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

Name	Description	Price
Bacteria glycerol stock	Bacteria glycerol stocks will be in sterile 50% glycerol in cryogenic tubes.	Inquire
Bacterial transformation/Miniprep/ Restriction digestion assay/PCR		Inquire
Customize virus packaging	1-Turnout time It may take 15day after receiving the DNA plasmid or after cloning is done 2-Quality control Confirmed QC DNA sequencing of target gene Titer- Titer of the virus tested by qPCR using standard. Titer- 10X¹¹- to 10X¹³ GC/ml Purity- AAV sample subject load on SDS PAGE followed by color staining. This assay confirms to see the AAV capsid protein VP1, VP2, and VP3 molecular weight and relative density. Endotoxin Free-To avoids the endotoxin-free AAV virus. We used an Endotoxin-free DNA extraction Kit (Qiagen). On-demand QC with or without extra charges Sterility- In this assay, the AAV virus treated to the in-vitro cell line and incubate for 2-4 days to suspect any bio-contamination. Transducibility-We can check in animal mice model in collaboration with Lobo Lab Electron Microscopy- NA	Inquire
DNA Prep Mini		Inquire
Endotoxin free DNA Maxi Prep	One DNA maxi prep reaction. Total amount of DNA may vary	Inquire
Lentivirus Packaging	Volume 1000ul, Ultrapurified in-vivo grade Titer- 10^8 to 10^9 GC/ml	Inquire
Lentivirus purified supernatant	10-20 ml purified supernatant lentivirus ~10^6 TU/ml	Internal $500.00 each External Academic Non-profit $1,000.00 each
Mutation or insertion or miRNA or big DNA or guide RNA cloning		Inquire
shRNA, guideRNA Screening or INDEL assay	1-shRNA Screening in cell line of your interest by qPCR or western blot. 2-gRNA cloning and screening of the guides by using qPCR, western or IHC. 3-INDEL assay by determination of mutations in your target gene by using Sanger sequencing.	Inquire
Stock Virus	1- Titer- 10X¹¹- to 10X¹³ GC/ml 2-Quality control Confirmed QC DNA sequencing of target gene Titer- Titer of the virus tested by qPCR using standard. Titer- 10X¹¹- to 10X¹³ GC/ml Purity- AAV sample subject load on SDS PAGE followed by color staining. This assay confirms to see the AAV capsid protein VP1, VP2, and VP3 molecular weight and relative density. Endotoxin Free-To avoids the endotoxin-free AAV virus. We used an Endotoxin-free DNA extraction Kit (Qiagen). On-demand QC with or without extra charges Sterility- In this assay, the AAV virus treated to the in-vitro cell line and incubate for 2-4 days to suspect any bio-contamination. Transducibility-We can check in animal mice model in collaboration with Lobo Lab Electron Microscopy- NA	Inquire

Search available services: View: by category alphabetically

Name	Description	Price
▼ ► Virus (2)
Customize virus packaging	1-Turnout time It may take 15day after receiving the DNA plasmid or after cloning is done 2-Quality control Confirmed QC DNA sequencing of target gene Titer- Titer of the virus tested by qPCR using standard. Titer- 10X¹¹- to 10X¹³ GC/ml Purity- AAV sample subject load on SDS PAGE followed by color staining. This assay confirms to see the AAV capsid protein VP1, VP2, and VP3 molecular weight and relative density. Endotoxin Free-To avoids the endotoxin-free AAV virus. We used an Endotoxin-free DNA extraction Kit (Qiagen). On-demand QC with or without extra charges Sterility- In this assay, the AAV virus treated to the in-vitro cell line and incubate for 2-4 days to suspect any bio-contamination. Transducibility-We can check in animal mice model in collaboration with Lobo Lab Electron Microscopy- NA	Inquire
Stock Virus	1- Titer- 10X¹¹- to 10X¹³ GC/ml 2-Quality control Confirmed QC DNA sequencing of target gene Titer- Titer of the virus tested by qPCR using standard. Titer- 10X¹¹- to 10X¹³ GC/ml Purity- AAV sample subject load on SDS PAGE followed by color staining. This assay confirms to see the AAV capsid protein VP1, VP2, and VP3 molecular weight and relative density. Endotoxin Free-To avoids the endotoxin-free AAV virus. We used an Endotoxin-free DNA extraction Kit (Qiagen). On-demand QC with or without extra charges Sterility- In this assay, the AAV virus treated to the in-vitro cell line and incubate for 2-4 days to suspect any bio-contamination. Transducibility-We can check in animal mice model in collaboration with Lobo Lab Electron Microscopy- NA	Inquire
▼ ► Cloning (2)
Name	Description	Price
shRNA, guideRNA Screening or INDEL assay	1-shRNA Screening in cell line of your interest by qPCR or western blot. 2-gRNA cloning and screening of the guides by using qPCR, western or IHC. 3-INDEL assay by determination of mutations in your target gene by using Sanger sequencing.	Inquire
Mutation or insertion or miRNA or big DNA or guide RNA cloning		Inquire
▼ ► DNA Prep (4)
Name	Description	Price
Bacteria glycerol stock	Bacteria glycerol stocks will be in sterile 50% glycerol in cryogenic tubes.	Inquire
DNA Prep Mini		Inquire
Endotoxin free DNA Maxi Prep	One DNA maxi prep reaction. Total amount of DNA may vary	Inquire
Bacterial transformation/Miniprep/ Restriction digestion assay/PCR		Inquire
▼ ► Lenti Virus production (2)
Name	Description	Price
Lentivirus Packaging	Volume 1000ul, Ultrapurified in-vivo grade Titer- 10^8 to 10^9 GC/ml	Inquire
Lentivirus purified supernatant	10-20 ml purified supernatant lentivirus ~10^6 TU/ml	Internal $500.00 each External Academic Non-profit $1,000.00 each