The facility is setup to cater for the production of lentiviruses and adeno-associated viruses.
The Viral Facility is a shared facility which provides the space, training, equipment and other resources to help in the production of replication-deficient viral vectors as well as in-vitro and in-vivo viral experiments.
The facility is built as a PC2 containment area and complies with Australian standards AS2982, AS2243 and AS2647. The facility is located in the Physiology department (North Wing of the Medical School Building) on the 5th level (Rooms N511, N512, N512A and N513).
There are four main areas within the facility:
- Anteroom (Rm N512A) is the main entry to the facility with an air-lock system containing a "gowning up" area prior to entering the labs.
- Cell Preparation Laboratory (Rm N513) which is used for the propagating cell lines and/or establishing primary cell cultures only.
- Viral Preparation Laboratory (Rm N512) which is used for the production, amplification, concentration and purification of viral vector particles.
- Viral Experimental Laboratory (Rm N511) which is used for in-vitro and in-vivo viral experiments.
Please contact Prof Andrew Allen or Angela Connelly, if you have any issues or questions regarding the Viral Facility.
Prof Andrew Allen
Ph: 61 3 8344 5838
Ph: 61 3 8344 5580
Viral Vector Overview
Replication-deficient viral vectors have been successfully used as tools for gene delivery by many laboratories around the world. Some of the advantages of using viral vectors over conventional gene delivery/pharmacological methods are:
- Broad or cell-specific targeting both in-vitro and in-vivo
- Infect dividing and non-dividing cells.
- Stable gene expression that can last for weeks to months (alternative to transgenic models)
- Use in many animal models (from rodents to primates).
- Diverse range of applications (from basic research to gene therapy).
Max. insert size
High titres, safe
Low titres, random
Specific integration (safe),
Small packaging capacity
All users will be charged based on the number of virus preps. Cost/viral prep* is as follows:
Lentivirus = $1280
AAV = $ 990
Additional charges for assistance based on the level of assistance required:
Full = $1075
Partial = $215
*The cost has been calculated to include equipment running servicing and running costs for the facility, including ultracentrifuge, biosafety cabinets, microscopes, Liquid nitrogen, C02, incubators, autoclave, etc. Also included is the average cost of consumables and reagents used for making a specific virus (large scale) according to protocols provided. Rates may also vary depending on changes to the prices of the items, or follow a different protocol that uses more or less of the listed items.
Prior to registering as a user, you must have:
In order to register as a user and access the facility, you must:
To make bookings go to Google Calendar.
List of Consumables/Reagents and equipment available
Ad.CMV.GFP (AdGo) –
ubiquitous cytomegalovirus promoter driving GFP
glial-specific enhanced GFAP promoter driving nuclear LacZ
neuron-specific enolase promoter driving LacZ
neuron-specific tubulin-α1 promoter driving EGFP
catecholamine neuron-specific synthetic dopamine β hydroxylase promoter driving LacZ
ubiquitous cytomegalovirus promoter driving EGFP
ubiquitin promoter driving EGFP
pan-neuronal calcium/calmodulin-dependent kinase II promoter driving EGFP
pan-neuronal human synapsin 1 promoter driving EGFP
pan-neuronal chicken β-actin promoter withcytomegalovirus (CMV) immediate-early enhancer driving tandom dimer tomato (red fluorescent) protein
catecholamine neuron-specific synthetic dopamine β hydroxylase promoter driving EGFP
serotinergic neuron-specific promoter driving EGFP
All are serotype 1/2 and contain chicken β-actin promoter with cytomegalovirus (CMV) immediate-early enhancer
(membrane bound EGFP)
(membrane bound tdTomato)
(endoplasmic reticulum-targeted tdTomato)
(pan-neuronal chicken β-actin promoter withcytomegalovirus (CMV) immediate-early enhancer driving Channel rhodopsin- mCherry)
Gateway Vector Conversion System (Invitrogen; cat no: 11826-029)
Multisite Gateway Pro2.0 (Invitrogen; cat no: 12537102)
Shuttle plasmids: |
Packaging plasmids (AAV2):
Restriction maps and sequences provided on request
HEK293FT (Invitrogen; cat no: R700-07)
AAV293 (Stratagene; cat no: 240073)
HEK293A (Invitrogen, catno: R70507)