# Viral Production and CRISPR Engineering

> **NIH NIH P30** · UNIVERSITY OF CALIFORNIA, SAN DIEGO · 2023 · $88,599

## Abstract

ABSTRACT
Viral Production and CRISPR Engineering Core
The NEI UCSD Center Core Grant for Vision Research Viral Production and CRISPR Engineering Core is
focused on the production of adeno-associated virus (AAV) and lentivirus (LV) vectors to allow for genome
engineering in vivo. Over the last several years, Shiley Eye Institute faculty have been heavily invested in
using genetic tools like high-throughput functional genomic screening (e.g., Wahlin and Welsbie) and
human genetics (e.g., Weinreb, Ayyagari, Borooah) to identify genes involved in various ophthalmic
diseases, including glaucoma, age-related macular degeneration (AMD) and inherited retinal degenerations
(IRDs). To be able to study the function of these genes in rodent models of disease, it is necessary to
produce gain-of-function (GOF) and loss-of-function (LOF) conditions. By fusing Cas9 to transcriptional
activators and repressors, technologies like CRISPR activation (CRISPRa) and inhibition (CRISPRi), as well
as conventional Cas9-mediated mutagenesis, can be used to produce the necessary GOF and LOF
conditions. Moreover, novel CRISPR technologies are being developed therapeutically to edit specific base
pairs or rewrite larger sections of genome using homology-directed repair (HDR). The key to all of these is
the viral delivery of genes encoding these large Cas9 proteins as well as optimized guide RNAs (gRNAs)
that direct Cas9. Finally, AAV can be used to express cDNAs (independent of CRISPRs). In all cases,
faculty are using a variety of outside cores for AAV production, resulting in high costs, variable quality and
only a limited selection of reagents. The Viral Production and CRISPR Engineering Core has access to
compact regulatory elements that can be used to create all-in-one AAV vectors that deliver Cas9 as well as
guide RNAs. The Core has a suspension cell culture setup, AAV and LV HEK293F producer cells, Cytiva
AKTA Pure fast protein liquid chromatography (FPLC) system and BioRad digital droplet PCR (ddPCR)
system for AAV/LV production, purification and titering. There is a dedicated part-time technician and
extensive know-how regarding AAV and CRISPR engineering that will be available to faculty.

## Key facts

- **NIH application ID:** 10709406
- **Project number:** 2P30EY022589-11
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN DIEGO
- **Principal Investigator:** Derek Stuart Welsbie
- **Activity code:** P30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $88,599
- **Award type:** 2
- **Project period:** 2023-09-01 → 2028-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10709406

## Citation

> US National Institutes of Health, RePORTER application 10709406, Viral Production and CRISPR Engineering (2P30EY022589-11). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10709406. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*