# Developing a CRISPR-based forward-genetic screening method in embryonic zebrafish

> **NIH NIH R15** · BRIGHAM YOUNG UNIVERSITY · 2024 · $449,350

## Abstract

Abstract
 One of the best methods for identifying novel genes and mutations affecting embryonic development is
forward genetic screening. However, these screens require a large number of animals to be screened to
identify a single novel mutant due to the inefficiency of chemical mutagenesis. Our current R15 finalized a
method for enzymatic generation of CRISPR libraries and used the method to generate sgRNAs targeting all
of, and only, the genes expressed in the developing zebrafish heart. The sgRNA templates were then divided
into eight groups of ten, transcribed, and injected into single-cell zebrafish embryos. Amazingly, all eight pools
showed clear morphological heart phenotypes. Thus, we are applying for this renewal grant to continue work
on the screen. We have identified a gene from the first round of the screen, sytl5. We will describe this gene’s
role in heart development, identify other mutated genes in our screen, and characterize their phenotypes in
detail. We also propose to optimize screen parameters to maximize our efficiency and to apply our CRISPR
library to a tissue-specific gene editing screen. In addition to the novel mutants identified in the screen, this
method will be able to be adapted to any tissue in any species, reducing the resources needed to conduct
forward genetic screens and increase the speed of gene discovery in a number of species and developmental
processes.

## Key facts

- **NIH application ID:** 10974321
- **Project number:** 2R15HD098969-02A1
- **Recipient organization:** BRIGHAM YOUNG UNIVERSITY
- **Principal Investigator:** JONATHON T HILL
- **Activity code:** R15 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $449,350
- **Award type:** 2
- **Project period:** 2019-08-01 → 2027-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10974321

## Citation

> US National Institutes of Health, RePORTER application 10974321, Developing a CRISPR-based forward-genetic screening method in embryonic zebrafish (2R15HD098969-02A1). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10974321. Licensed CC0.

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