Vector Development and Production Core
Director: Bakhos Tannous
Location: Building 149 13th Street, Room 6309, Charlestown
| Core Summary: |
The Vector Development and Production Core provides viral vectors with custom-designed promoters and reporter genes and capacity for gene regulation.
| Personnel/Contact Information: |
Director: Bakhos Tannous, Ph.D.
Phone: (617) 726-6026
Fax: (617) 724-1537
Email: btannous@partners.org
Other Staff:
Research Technician: Danielle Morse
Phone: (617) 726-6027
Email: dmorse2@partners.org
| Facilities: |
Location of the Core:
The facility is located at Building 149 13th Street, Room 6309, Charlestown.
| Services: |
This Core will advise investigators on lentivirus, retrovirus and adeno-associated virus (AAV) vectors most appropriate for their research needs based on relative infectivity of cells of interest, use for culture or in vivo studies, promoter considerations, hypotheses being addressed, and appropriate reporter genes. The Core will provide appropriate lentivirus, retrovirus and AAV plasmid backbones and construction information and assistance to investigators, as well as packaging vectors for them.
Investigators using the lentivirus vector will be supplied with a plasmid backbone carrying multiple cloning sites (MCS) downstream of a cytomegalovirus (CMV) IE promoter. Other plasmids are also available with a reporter gene such as GFP, RFP under an IRES element. These vectors will express both the transgene of interest as well as the reporter gene.
For investigators interested in retrovirus vectors, a number of constructs are provided, including drug selection cassettes for puromycin, zeomycin and neomycin, as well as reporter genes, eGFP or luciferase, and cloning sites for transgenes of interest under the 5' LTR promoter.
AAV vectors made available through the core will carry AAV2 ITR elements serotyped with either AAV1, 2, 5, 8, 9, or rh10 capsids. A number of tissue-specific promoters are available together with the constitutively active CBA promoter. A bicistronic vector expressing the transgene as well as GFP is available to facilitate identification of vector-transduced cells in cell culture and in vivo. Basic vectors encoding EGFP, firefly luciferase (Fluc), Gaussia luciferase (Gluc), and Cre recombinase (under CBA or Synapsin-1 promoters) are available.
Investigators will be responsible for inserting their genes of interest into the MCS, validating correct construction by restriction analysis, testing bioactivity by transfection assays (when possible), and supplying clean maxi-prepped DNA for packaging.
The Core will maintain high quality stocks of packaging plasmids and will carry out packaging and “purification”, titering and allocation of vectors for the investigators.
| Getting Started: |
To access the core, please contact Bakhos Tannous at (617) 726-6026 or by email at
btannous@partners.org.
| Departmental Web Link: |
http://www2.massgeneral.org/ncs/neuro_core_VectorDevelopmentandProduction.htm
