# Phenotyping novel organogenesis lethal KOMP alleles

> **NIH NIH R01** · UNIVERSITY OF MASSACHUSETTS AMHERST · 2022 · $410,110

## Abstract

Project Summary.
Although the mammalian genome has been sequenced for more than 15 years numerous gaps lie in
understanding how these sequences function in the context of the organism. The goal of the knock-out mouse
project (KOMP) is to functionally annotate the coding sequences of the genome by knocking out individual
genes and assessing their role in the mouse. An overarching goal of the project is to determine the role of
individual genes in a plethora of complex traits that underlie diseases that pose significant risks to human
health including heart disease, diabetes, metabolic syndrome, cognition and high blood pressure. Importantly
over 25% of genes knocked out result in embryonic lethality. I currently collaborate on an existing KOMP
project designed to examine novel null alleles that are lethal prior to E9.5 and of the 88 examine and only 4
exhibit post-gastrulation defects. The goal of this project is to more effectively use our laboratories expertise in
post-gastrulation/early organogenesis to determine the primary defects of 25-40 novel KOMP-derived strains
that are critical during these developmental stages. We propose to analyze mutant strains that are present at
E9.5 but lethal by E12.5. In Aim 1a, we propose to identify the normal expression pattern of the deleted gene,
using either the LacZ reporter allele or in-house generated in situ probes. We will characterize each strain by
examining the onset of histological defects in the null embryos/extra-embryonic tissues compared with stage
matched controls, and by performing this analysis one day later, when we expect the early phenotype to be
more pronounced. In Aim 1b, we will perform a standard 4-color immunofluorescence on null and control
embryos, that is designed to detect defects in each of the 3 embryonic germ layers. We anticipate numerous
embryonic and extra-embryonic phenotypes and explain how particular phenotypes will be further examined
using existing resources in our laboratory. Finally, because we expect that many of the lines examined in Aim
1a and 1b will display primary defects in extra-embryonic tissues, we propose to further investigate the role of
3-5 of these genes in Aim1c by utilizing KOMP-generated conditional ready ES cells to delete that gene in the
embryo but retain expression in the extra-embryonic tissues. These conditional experiments, when compared
with the null experiments performed in Aim1a and 1b, will allow us to parse out the role of each novel gene in
the embryo-proper and in the extra-embryonic tissues. Together these analysis will not only assist with the
functional annotation of critical novel genes but will also lead to a better understanding of the etiology of
human pediatric diseases and will serve to widen the drug discovery pipeline.

## Key facts

- **NIH application ID:** 10397128
- **Project number:** 5R01HD096073-04
- **Recipient organization:** UNIVERSITY OF MASSACHUSETTS AMHERST
- **Principal Investigator:** KIMBERLY D TREMBLAY
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $410,110
- **Award type:** 5
- **Project period:** 2019-09-10 → 2024-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10397128

## Citation

> US National Institutes of Health, RePORTER application 10397128, Phenotyping novel organogenesis lethal KOMP alleles (5R01HD096073-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10397128. Licensed CC0.

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