Lentiviral Vector Viral Particles Production Core Platform
We offer comprehensive services in tissue culture and lentiviral vector production for University of Manitoba investigators, as well as provincial and international collaborators.
In-stock viral preparations
The core will maintain stocks of 293T cells, packaging and envelope plasmids (for lentiviral vector production), and control lentiviral vector plasmids (such as irrelevant shRNA silencing specificity control, scrambled shRNA silencing specificity control, GAPDH-silencing positive control) and an EGFP-expressing lentiviral vector (for evaluation of transduction efficiency in the cell type of interest). It will provide these reporter or control viral preparations to users at cost-recovery prices.
Custom-made viral preparations using the lentiviral vector plasmid provided by users
Using the standard lentiviral constructs, the Core will produce high quality lentiviral viral preparations in unconcentrated format (titer estimated to be ~1X106 TU/ml; one volume size) and concentrated format (titer estimated to be ~1X108 TU/ml; in small, medium and large sizes). We can package also lentiviral vectors of other sources; however, as lentiviral vector designs will affect packaging efficiency, the titer will need to be determined.
Significant impact of the platform
- To enhance research productivity by saving time that a researcher needs to prepare these viral preparations routinely.
- To facilitate the use of this vastly popular technique in basic and translational research. It will encourage new users who hesitate to set this up in the laboratory due to biosafety concerns, infrastructure requirements or high cost of purchasing small amount of virus to test from the commercial resources.
- To make full use of the unique resource of the lentiviral shRNA libraries we already have on campus. The two platforms will integrate and further enhance the competitiveness of the research projects/proposals of our researchers at the university.
What we offer
We suggested the use of concentrated Cppt2E (EGFP-expressing lentiviral vector) virus in testing transduction efficiency and in the optimizations of your cell type of interest. The use of concentrated Cppt2E virus (in stock item) in a titration experiment will also allow users to evaluate whether your cell type of interest requires the use of concentrated or unconcentrated virus format in your transduction experiments. BSL2 facility is required in the transduction of cells. An update of the biological inventory to include the use and storage of the lentiviral vectors in the users’ laboratory biosafety permit is required.
I'm not sure where to place this, does it belong on this page or should it go on the Services page? - CM
The Core consists of BSL-2 tissue culture facility and the ultracentrifugue set-up. These are housed on the fourth floor of the Apotex Building.
The four shRNA libraries are housed at System Biology laboratory at Manitoba Centre for Proteomics and Systems Biology, located within the Health Sciences Centre.
To avoid any possible contamination and maintain a clean laboratory environment, we recommend following some standard safety practices.
- Wear double gloves, face protection, and lab coat at all times
- Perform work in a limited access area in a Biological Safety Cabinet Class II
- Post a biohazardous warning sign
- Minimize splashes or aerosols with careful pipetting
- Refrain from using needles, blades and other sharp items
- Decontaminate work surfaces at least once a day and after any spill of viable material
- Decontaminate all cultures, stocks and other biological wastes before disposal using approved decontamination methods
- Transport biological materials from the laboratory in a sealed, durable, leak-proof container
Lentiviral Core Facility
750 McDermot Avenue West
University of Manitoba (Bannatyne campus)
Winnipeg, MB R3E 0T5 Canada