# Core D:  Mouse Perturbation Core

> **NIH NIH U19** · HARVARD MEDICAL SCHOOL · 2021 · $812,789

## Abstract

Mouse Perturbation Core D is central the mission of this U19 Program because it will execute
CRISPR/Cas9 forward genetic screens in mice to enable U19 investigators to identify novel regulators
of innate and adaptive immune responses to acute infection in mouse models. Core D will make an
innovative technology for conducting Cas9/CRISPR-mediated genetic screens easily accessible to all
U19 investigators. Core D will assist U19 investigators with screen design; generate and validate bone
marrow chimeric mice carrying curated pools of sgRNA; infect these bone marrow chimeras or
recipients of T cells carrying sgRNAs from these bone marrow chimeras with virus for U19
investigators; and assist U19 investigators with validation of candidate regulators in acute viral infection
models. Core D has expertise with all of these procedures and demonstrated success with using these
approaches to conduct forward genetic screens in vivo. To achieve these goals, the Specific Aims of
Core D are: 1) To generate and validate bone marrow chimeric mice carrying sgRNA libraries. Core D
will provide advice on screen design and then transduce the appropriate Cas9-expressing
hematopoietic progenitor cells with lentivirus carrying curated libraries of sgRNAs generated by Core C;
2) To infect bone marrow chimeric mice or recipients of T cells from these mice with virus. Core D will
infect bone marrow chimeras or recipients of transduced T cells from bone marrow chimeras for in vivo
screens (using LCMV Armstrong), and for in vivo validation studies of candidate regulators of innate
and adaptive immune responses to acute infection (using LCMV Armstrong or influenza virus). Core D
also will provide transduced DCs from bone marrow chimeras for ex vivo screens to identify regulators
of DC interactions with pathogens, pathogen components and T cells; 3) To generate, maintain and
validate TCR transgenic and immune-lineage-specific Cas9 expressing mice, and LCMV Armstrong
and influenza viral stocks. Core D will serve as a repository for Cas9-expressing mouse strains and
viral stocks. This centralized approach will standardize the set of procedures needed to conduct
forward genetic screens in vivo. Provision of this novel technology by a core provides an efficient and
cost-effective means to conduct these in vivo screens, to ensure uniformity and success in use of these
approaches by U19 investigators, and to facilitate comparisons of data across the U19 Program. Core
D will work closely with investigators in Project 1 and 2 to help them conduct in vivo screens and
validate candidate regulators, and with Core C for lentiviral pools of sgRNA for the screens, Core A to
facilitate communication and administrative oversight of Core D activities, and Core B through the
Project investigators for computational analyses of single cell data from immune populations.

## Key facts

- **NIH application ID:** 10207348
- **Project number:** 5U19AI133524-05
- **Recipient organization:** HARVARD MEDICAL SCHOOL
- **Principal Investigator:** Arlene H. Sharpe
- **Activity code:** U19 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $812,789
- **Award type:** 5
- **Project period:** 2017-07-05 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10207348

## Citation

> US National Institutes of Health, RePORTER application 10207348, Core D:  Mouse Perturbation Core (5U19AI133524-05). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10207348. Licensed CC0.

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