# Genome-wide CRISPR-Cas9 screens in insect cells to characterize insecticidal toxins

> **NIH NIH R01** · BOSTON CHILDREN'S HOSPITAL · 2024 · $794,860

## Abstract

Project Summary
 Genome-wide screens using CRISPR-Cas9 technology have revolutionized studies of
host-pathogen interactions, leading to identification of many key host cellular factors required for
the actions of human pathogens and bacterial toxins. However, this powerful approach has yet to
be utilized in insect cells to uncover host factors required for insect-borne pathogens, which are
responsible for a long list of infectious diseases such as malaria, Dengue, West Nile, Zika, and
Lyme diseases. This is largely due to a technical barrier: the inability to efficiently deliver genome-
wide guide RNA library into insect cells. We recently overcame this barrier and developed a
genome-wide CRISPR-Cas9 screening method in cultured Drosophila cells. Using this method,
and by leveraging the expertise of Dr. Norbert Perrimon’s lab in insect models and of Dr. Min
Dong’s lab in bacterial toxins, we carried out extensive preliminary studies, leading to the
identification of a potential insect receptor for a member of the major bacterial toxin family known
as Tc toxins, demonstrating the power and feasibility of our unbiased genome-wide screen
approach. Building on these successes, in Aim 1 we will focus on further development and
validation of genome-wide screens with major Tc toxin family members to establish a mechanistic
understanding of toxin-receptor interactions and their role in pathogenesis in vitro and in vivo. We
further propose to expand our approach to establish the first genome-wide CRISPR-Cas9
screening method and tools in mosquito cells in Aim2, and then utilize this approach to identify
key host factors for two novel bacterial toxins that showed selective toxicity on mosquito but not
Drosophila cells. The success of our proposal will uncover receptors and key host cellular factors
for important bacterial toxins and establish generalizable methods and essential tools for
investigating pathogens and toxins at genome-wide scale in insect cells relevant to transmitting
human infectious diseases.

## Key facts

- **NIH application ID:** 10843913
- **Project number:** 5R01AI170835-03
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **Principal Investigator:** Min Dong
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $794,860
- **Award type:** 5
- **Project period:** 2022-06-15 → 2025-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10843913

## Citation

> US National Institutes of Health, RePORTER application 10843913, Genome-wide CRISPR-Cas9 screens in insect cells to characterize insecticidal toxins (5R01AI170835-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10843913. Licensed CC0.

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