# Bacterial Genetics Core D

> **NIH NIH U19** · BRIGHAM AND WOMEN'S HOSPITAL · 2021 · $402,576

## Abstract

Core D. Bacterial Genetics Core
Core Leader: Jeremy Rock
ABSTRACT
Stewart Cole and colleagues determined the complete genome sequence of Mycobacterium tuberculosis (Mtb)
in 1998 (1). This landmark achievement heralded a new age in mycobacterial research, including the
development of organism-wide gene knockout and knockdown technologies that made it possible to determine
the roles of specific mycobacterial genes in survival and host response. However, despite the ability to
interrogate thousands of new potential targets, few new genes have advanced as targets for active clinical
drug development. This shortfall stems, in part, from key technical limitations in the ability to systematically
interrogate the Mtb genome on an organism-wide basis. To help overcome this limitation, Jeremy Rock and
colleagues developed CRISPRi interference (CRISPRi) technologies that achieve robust, programmable gene
silencing in Mtb. The Rock laboratory has now validated a genome-scale library of 96,700 independent
CRISPRi mutants, which comprise the central technology of the Bacterial Genetics Core D. These efforts have
resulted refined CRISPRi design rules, allowing the generation of highly efficacious and specific CRISPRi
knockdown for nearly all Mtb genes, including methods for titratable knockdown of essential genes. The
Bacterial Genetics Core will support Project 1 by designing and constructing individual and pools of Mtb
CRISPRi mutants to identify new lipids that are downstream of genes involved in virulence, barrier function and
Mtb strain variations in human patients. Core D will support Project 2 by providing genetic mutants within the
MtrAB signal transduction pathway, a central mediator of intrinsic multi-drug resistance in Mtb. In addition,
CRISPRi will be used to silence genes involved in the mycobacterial drug response, intrinsic drug resistance to
rifampicin, as well as Mtb envelope composition. These studies will identify novel genetic and biochemical
targets for development of new anti-mycobacterial drugs. Further, these experiments inform strategies for
augmenting the efficacy of existing drugs through targeting bacterial genes that modulate the sensitivity of Mtb
to antibiotics.

## Key facts

- **NIH application ID:** 10271482
- **Project number:** 1U19AI162584-01
- **Recipient organization:** BRIGHAM AND WOMEN'S HOSPITAL
- **Principal Investigator:** Jeremy Michael Rock
- **Activity code:** U19 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $402,576
- **Award type:** 1
- **Project period:** 2021-07-01 → 2026-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10271482

## Citation

> US National Institutes of Health, RePORTER application 10271482, Bacterial Genetics Core D (1U19AI162584-01). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10271482. Licensed CC0.

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