The GCD core provides technical support for the production of recombinant viral vectors, and facilitate targeted delivery of genes, cells and other reagents to ocular tissues.

Viral vector design service: 

The core director and staff will provide consultation regarding viral vector choice and design, as well as available common vector backbones and packaging helpers from the core (see Table 1).


Table 1. Currently Available Viral Repository in the GCD Core

 

Vector System Packaging Helpers Promoter-Transgenes

Lentiviral vectors

(3rd generation, SIN vector)

pLP.VSVG
pLP1
pLP2

CVM-EGFP
CMV-CNTF
CMV/Myo-Myo7a
Ubi-rTA
TetO-FUW-OSKM
TetO-EGFP
TeTo-Ngn2-EGFP-PuryR
LentiCRISPRv2GFP

AAV vectors pHelper
pRepCap (serotype 2,5,8)
pCap.Shh10
pCap.7m8
pAAV-CAG-GFP
pAAV-CAG-Cre
pAAV-DFAP-Cre
PaaV-CMV-SaCas9;U6-sgRNA
pAAV-CAG-STAT3
pAAV-Tet-RFP;Ubi-rTA

Viral Production Laboratory:  

This biosafety level 2 (BSL2) facility is equipped with biosafety cabinet, TC incubators, inverted microscope, centrifuges, and other necessary equipment for molecular biology operation, recombinant lentivirus and AAV production and concentration.  The viral production laboratory will provide common cell lines, viral vectors, and packaging helpers DNAs.  High titer lentiviruses and small test batch of AAVs can be produced by this laboratory with investigator-provided specific viral vectors.

 

Biosafety cabinet for viral production

Biosafety Cabinet for Viral Production

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Intraocular injection facility:  

This his biosafety level 2 (BSL2) facility can house up to 112-cage special isolator cages.  The same facility has a laminar flow hood outfitted with a Leica stereomicroscope with LED light source for eye injections.  The facility has Shutter glass pipette puller and FemtoJet pressure-operated micromanipulator with microloader for intraocular injections for investigators with appropriate institutional approvals.

Biosafety cabinet for eye injection

Biosafety Cabinet for Eye Injection

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Special isolator cage rack

Special Isolator Cage Rack

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BTX Electroporator

BTX Electroporator

FemtoJet Microinjector

FemtoJet Microinjector

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Leica Stereomicroscope

Leica Stereomicroscope

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