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Stem Cell Gene Transfer and Viral Vector Core (SCGTC)

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

 

LVV-CMV-X-Pgk-Puro

LVV-CMV-GFP-PGK-Puro

LZRS-Ires-GFP

 

LZRS-MCS

PLK01-ScrmshRNA-Puro

LZRS-RFP

 

LZRS-X-Ires-Puro

PLK01-ScramshRNA-Neo

LZRS-YFP

 

LZRS-X-Ires-GFP

 

LZRS-LZ

 

 

 

LZRS-Cre

 

 

 

LZRS-Cre-Ires-puro

 

 

 

LZRS-Cre-Ires-GFP