# Core C - Virus/Vector

> **NIH NIH P01** · UNIVERSITY OF WISCONSIN-MADISON · 2024 · $78,206

## Abstract

CORE C – PROJECT SUMMARY/ABSTRACT
Core C. Virus/Vector Core
Eric Johannsen, Core Leader
Core C (Virus Core) provides time and cost-efficient generation, validation, distribution, and archival storage of
wild-type (WT) and mutant viruses needed by all 6 research groups of this Program Project for their proposed
studies. The Specific Aims are: (1) to produce validated stocks of WT, mutant, and revertant Epstein-Barr virus
(EBV) and Kaposi Sarcoma Herpesvirus (KSHV); (2) to produce validated stocks of infectious mouse
papillomavirus (MmuPV1) and human papillomavirus (HPV) pseudoviruses; and (3) to produce validated
lentivirus library stocks. Mutant EBV (for Projects 2, 3, and 4) or KSHV (for Project 2) will be generated in E. coli
starting with appropriate BACs using the scarless En Passant method.1 We can also make mutants, when
appropriate in eukaryotic cells using CRISPR/Cas9 editing. In addition to standard methods, we have established
a bioinformatic pipeline to analyze next generation sequencing data of herpesvirus genomes to ensure there are
no unintended mutations within their unique regions. Whenever necessary, phenotypes will be validated using
trans-complementation or the construction of revertant EBV/KSHV genomes. Virus stocks of WT, mutant, and
revertant variants of EBV or KSHV will be generated using a protocol developed by Dr. Sugden, titered, and
stored for use by all three EBV groups. Papillomavirus virions and pseudovirions will be generated (for Project
1) by co-transfection of 293T cells with (i) the desired capsid protein-expression plasmids, and (ii) the desired
viral or luciferase/GFP-encoding DNAs targeted for encapsidation using protocols previously published by the
Lambert/Ahlquist laboratories. Lentivirus libraries will be packaged and tittered using standardized protocols for
use in Projects 2 and 4. Lenti and retrovirus transduction vectors with different antibiotic resistance genes have
been generated and will be provided to Projects to allow transduction of multiple genes into cells. In addition to
increased cost efficiency and quality control, the existence of this Core facilitates the use of common virus stocks
that facilitate the interpretation of complementary data generated among the various research groups to foster
shared aims within the projects. All 6 research groups associated with the 4 projects will be served by this Core
facility.

## Key facts

- **NIH application ID:** 10837993
- **Project number:** 2P01CA022443-46A1
- **Recipient organization:** UNIVERSITY OF WISCONSIN-MADISON
- **Principal Investigator:** ERIC C. JOHANNSEN
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $78,206
- **Award type:** 2
- **Project period:** 1997-02-01 → 2029-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10837993

## Citation

> US National Institutes of Health, RePORTER application 10837993, Core C - Virus/Vector (2P01CA022443-46A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10837993. Licensed CC0.

---

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