# Phenotype-driven approach to understanding the function of craniofacial regulators using IMPC-generated mouse strains

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · 2022 · $103,471

## Abstract

ABSTRACT
Many of the datasets resulting from genome-wide approaches lack functional validation in living organisms.
While the laboratory mouse is often used as an experimental model, a large number of mouse genes have
unknown functions. The International Mouse Phenotyping Consortium (IMPC) is building the first catalogue of
mammalian genome function by generating knockout (KO) mouse strains for every protein-coding gene in the
genome. Taking advantage of this opportunity, the two co-PIs have designed this proposal in response to
NIH PAR-17-005 for phenotyping IMPC embryonic and perinatal lethal KO mouse lines. Our focus is on
mutations that affect the craniofacial complex, based on our expertise in modeling craniofacial malformations in
the mouse. While craniofacial defects represent one third of all human birth defects, our knowledge of the
underlying cellular and molecular mechanisms remains poor. To select mutant mouse lines for in-depth
phenotyping of craniofacial abnormalities, we generated algorithms to intersect the current list of IMPC lethal
/subviable KO strains exhibiting craniofacial defects with: 1) all genes present in transcriptomes of mouse
embryonic craniofacial domains that were either generated in our labs or available in the FaceBase database;
and 2) all significant ChIP-Seq peaks for binding of the enhancer-associated protein p300 in mouse embryonic
whole faces present in the FaceBase database. By searching the Mouse Genome Informatics (MGI) database
for phenotypic data and by prioritizing genes that have unknown or poorly defined roles in craniofacial develop-
ment, we restricted the number of chosen genes to N=30. These comprise regulators of various cellular
functions that cause embryonic lethality and craniofacial defects when disrupted in the mouse. Preliminary
phenotyping of the KO mouse line for Zfhx4, a gene that exhibited highest maxillary enrichment in our RNA-
Seq dataset, revealed that all Zfhx4 KO embryos present cleft palate and maxillary hypoplasia, providing proof
of concept for the effectiveness of the proposed strategy. Accordingly, we will characterize 30 mutant lines (15
per lab over 5 years) via the following specific aims: AIM 1: Examination and classification of 30 IMPC
mutant mouse lines for craniofacial phenotypes. Employing a two-stage phenotyping pipeline, we will
categorize the craniofacial defects for each line and narrow the time of onset. AIM 2. Deep characterization
of craniofacial phenotypes. Based on three phenotyping platforms, we will uncover the function(s) of the 30
chosen genes in craniofacial development through in-depth analyses. Platform A will characterize early
craniofacial anomalies including defects in branchial arch patterning, as well as primary palate morphogenesis
and fusion (E8.5-11.5); Platform B will characterize later abnormalities of secondary palate fusion (E11.5-15.5);
and Platform C will dissect perturbations of craniofacial shape/morphology and skull ossification. She...

## Key facts

- **NIH application ID:** 10589996
- **Project number:** 3R01DE028753-04S1
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
- **Principal Investigator:** Jeffrey Ohmann Bush
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $103,471
- **Award type:** 3
- **Project period:** 2019-04-01 → 2024-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10589996

## Citation

> US National Institutes of Health, RePORTER application 10589996, Phenotype-driven approach to understanding the function of craniofacial regulators using IMPC-generated mouse strains (3R01DE028753-04S1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10589996. Licensed CC0.

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