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
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
Gene cloning
5µg Vector in water
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
$900.00
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
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
Welcome!
