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.
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 |
| 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 |
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
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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
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Special isolator cage rack
BTX Electroporator
FemtoJet Micorinjector
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Leica Stereomicroscope
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injection facility
