# Molecular Virology Core

> **NIH NIH P01** · INDIANA UNIVERSITY INDIANAPOLIS · 2024 · $348,023

## Abstract

PROJECT SUMMARY/ABSTRACT
To understand the critical factors that negatively impact gene therapy for hemophilia A, our program will
investigate the key problems associated with AAV vector production and design, factor VIII (FVIII) expression,
and role of the immune system. Proposed gene transfer studies critically rely on strict control of vector
composition and thorough characterization of vector properties before and after vector administration.
Unfortunately, in-depth characterization of vectors is stymied by the lack of knowledge of some of the critical
parameters of a given vector preparation that may be key to vector performance. This is exemplified by
challenges in analyzing the impurities within AAV vector products such as defective viral particles that closely
resemble the vector itself. The scientists of the proposed Molecular Virology Core (MVC) developed a series of
new assays on AAV vectors which provide additional critical parameters to be analyzed for safety of gene
therapy. This unprecedented depth of analysis will provide Projects 1, 2, and 3 with the knowledge of vector
composition that they require to interpret the outcomes of gene transfer experiments and help develop safer
vectors. The MVC will pursue the following Specific Aims: 1) Manufacturing of quality vectors to support all
projects: For results between studies and projects to be comparable, identical standards of vector production
need to be applied. The proposed manufacturing process will further eliminate variables, e.g., use of plasmid
backbone-free production, among other innovations. In addition, MVC will aid Projects 1, 2 and 3 in vector
development, incorporating new discoveries and generating novel AAV vectors that improve hepatic FVIII gene
delivery. 2) Providing in-depth molecular characterization of AAV vectors for in vitro and in vivo studies:
MVC will provide single AAV genome analysis using a newly developed assay based on PacBio sequencing and
bioinformatic analysis of the sequence data. Traditional AAV vector assays such as AAV titering by ELISA,
Southern blot analysis, silver staining as well as digital PCR/qPCR will also be performed. 3) In-depth analysis
for AAV genome status post-delivery. AAV genomes status in vivo will be analyzed by next generation
sequencing as well as traditional Southern blot and qPCR. AAV transcriptome analysis will also be included.

## Key facts

- **NIH application ID:** 10778204
- **Project number:** 5P01HL160472-03
- **Recipient organization:** INDIANA UNIVERSITY INDIANAPOLIS
- **Principal Investigator:** WEIDONG XIAO
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $348,023
- **Award type:** 5
- **Project period:** 2022-02-05 → 2027-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10778204

## Citation

> US National Institutes of Health, RePORTER application 10778204, Molecular Virology Core (5P01HL160472-03). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10778204. Licensed CC0.

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