Stem Cell Gene Transfer and Viral Vector Core (SCGTC)
Scientific Director: Dr. S. Ghazizadeh, Department of Oral Biology and Pathology
Phone: 631-632-3138
Email: Soosan.Ghazizadeh@stonybrookmedicine.edu
The generation of high quality recombinant viral vectors is complex and requires proper safety training and institutional approvals and therefore not easily adapted by new investigators. The SCGTC will provide this critical support to assist investigators in the genetic modulation of stem cells as a means of studying gene function. Standard retroviral and lentiviral vectors have been successfully used to stably express or suppress genes in stem cells, and will be the primary vehicles for transduction. Additional core competencies in adenoviral and adeno-associated virus (AAV) generation and also available, and provided in conjunction with Dr. Patrick Hearing (Microbiology and Molecular Genetics).
Key functions of the SCGTC Core will be to:
(1) Provide purified viral expressing genetic cocktails for generation of iPSCs.
(2) Facilitate the design, construction and propagation of retro/lentiviral vectors for gene over expression in stem cells.
(3) Facilitate the design, construction, propagation, and purification of adenovirus and AAV
(4) Generate and characterize retro/lentiviral vectors for gene-restricted knockdowns using shRNAs or miRNAs
(5) Gene editing technology using Crisper/Cas (Dr. ZongDong Li, Dept. Medicine)
(5) Education and training of investigators and trainees in viral vector design and manipulation.
The SCGTC will provide guidance on the generation of particular shRNAs, cDNAs, and/or microRNAs and individual investigators will test these for genetic modulation and identify at least two that work efficiently. All viruses provided by the SCGTC will be sequenced for verification of targeting constructs prior to viral production and confirmation of stem cell transduction. For all studies, the SCGTC will maintain logs for all viral constructs, freeze viral aliquots for future need, and cryopreserve genetically modified cells for storage and distribution to other investigators.
Stringent quality control is implemented to exclude the presence of Mycoplasma and replication competent contaminating virus
Currently Available Viral Aliquots
Lentiviruses |
Retroviruses |
Adenoviruses |
Vector Backbone |
LVV-CMV-GFP |
LZRS-flg-hOCT4 |
Ad5-GFP |
PLK01-Puro |
LVV-CMV-RFP |
LZRS-flg-KLF4 |
Ad5-LZ |
PLK01-Neo |
LVV-CMV-LZ |
LZRS-flg-Sox2 |
Ad5-Cre-GFP |
pLV-EF1a-miRNA-puro |
LVV-EF1a-GFP |
LZRS-flg-cMyc |
|
LVV-EF1a- X–Ires-puro |
LVV-EF1a-GFP-PGK-Puro |
LZRS-GFP |
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LVV-CMV-X-Pgk-Puro |
LVV-CMV-GFP-PGK-Puro |
LZRS-Ires-GFP |
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LZRS-MCS |
PLK01-ScrmshRNA-Puro |
LZRS-RFP |
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LZRS-X-Ires-Puro |
PLK01-ScramshRNA-Neo |
LZRS-YFP |
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LZRS-X-Ires-GFP |
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LZRS-LZ |
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LZRS-Cre |
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LZRS-Cre-Ires-puro |
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LZRS-Cre-Ires-GFP |
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