# Core B Gene Perturbation Core

> **NIH NIH P01** · YALE UNIVERSITY · 2024 · $273,970

## Abstract

Core B –Gene Perturbation (Sharpe/Kuchroo):
This Core will integrate transgenic, knockout, and CRISPR/Cas9 gene perturbation technologies into the study
of TIGIT/CD155 and PD-1/PD-L1 functions and interactions. Core B has the expertise with both conventional
and conditional transgenic and knockout technology, as well as CRISPR/Cas9 engineering, which will enable
the generation of novel mouse strains for analyzing the functions of TIGIT/CD155 and PD-1/PD-L1 functions and
interactions in vivo. The Mouse Engineering Core provides a critical means by which the PPG will achieve its
goals of understanding how TIGIT/CD155 and PD-1/PD-L1 mediate their immunoregulatory functions.
Core B will provide PPG investigators with an important and unique collection of mouse strains for their studies.
To facilitate studies of the function of TIGIT and PD-1 in specific cell types, the Core will cross TIGIT (TIGITfl/fl)
and PD-1 (PD-1fl/fl) floxed mice with different Cre driver strains. Likewise, to enable analyses of the function of
CD155 and PD-L1 in specific cell types, the Core will cross CD155 (CD155fl/fl) and PD-L1 (PD-L1fl/fl) floxed mice
with different Cre driver strains. The initial priority will be to breed CD155fl/fl mice with PD-L1fl/fl and CD11cCre
and Zbtb46Cre mice to delete CD155 and PD-L1 in dendritic cells (DC). These strains will enable studies of the
role of CD155 and PD-L1, individually and together on DC. Core B also will generate CD155 mutant mouse
strains to support studies of CD155 cell intrinsic functions. The generation of CD155 ITIM mutant and CD155
cytoplasmic tail-less mouse strains to enable studies of bidirectional interaction of TIGIT with CD155 will be the
initial priority. Importantly, these CD155 mutations will preserve CD155 receptor binding and adhesion functions,
but alter intracellular signaling, thus enabling study of how these mutations impact CD155 cell intrinsic functions.
In addition, Core B will enable PPG investigators to identify mediators of TIGIT/CD155 and CD155/PD-L1
functions, individually and in combination, in effector and regulatory T cells. Core B will generate lentiviruses
with gRNAs for use in vivo in the CHIME (CHimeric IMmune Editing) CRISPR/Cas9 gene perturbation platform.
CHIME enables perturbation of genes in naïve lymphocytes, myeloid cells, and dendritic cells in vivo without
altering their development. Core B will use CHIME to enable studies to validate prioritized genes identified by
proteomic and transcriptional data in Project 1 (Kuchroo), Project 2 and Project 3 (Hafler). Core C will integrate
the data sets from the projects and assist with gene prioritization.
Finally, Core B will serve as a repository for new mouse strains generated during the course of this PPG. The
Mouse Engineering Core will provide breeding pairs of these strains to PPG investigators as needed and
cryopreserve strains.

## Key facts

- **NIH application ID:** 10839293
- **Project number:** 5P01AI039671-26
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Arlene H. Sharpe
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $273,970
- **Award type:** 5
- **Project period:** 1997-09-01 → 2028-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10839293

## Citation

> US National Institutes of Health, RePORTER application 10839293, Core B Gene Perturbation Core (5P01AI039671-26). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10839293. Licensed CC0.

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