Investigating Novel Mutations and Developmental Timing of Congenital Diaphragmatic Hernia

Ameleen Wong

Document Type

Article

Publication Date

8-9-2024

Keywords

JMG

JAX Location

In: Student Reports, Summer 2024, The Jackson Laboratory

Abstract

Congenital Diaphragmatic Hernia (CDH) is a mammalian-specific muscle critical for respiration and division of the thoracic and abdominal body cavities. CDH affects 1:3000 live births with a mortality rate of 30%, characterized by muscle weakness/partial loss of diaphragm muscle accompanied by lung hypoplasia. Development of CDH is primarily driven by genetic mutations, however, the total number of CDH critical genes remains a question in the field. This research presents a method to screen CDH patient mutations in mice, demonstrated by investigation of two novel gene mutations—Wt1 and Kalrn. To introduce mutations into fertilized mouse zygotes (E0.5 embryos) CRISPR/Cas9 technology was used. Edited E0.5 are transplanted into donor mice to allow for embryonic development past the stage of diaphragm patterning. Edited embryos were then analyzed for structural defects using X-Ray Micro-Computed Tomography to assess diaphragmatic integrity and lung development. The experimental results showed CDH-like phenotypes in mutant mouse embryos and further understanding of the roles of Wt1 and Kalrn mutations in CDH. We also were able to have a better understanding when herniations arose within Cdc42bpb from embryonic days E13.5 -E16.5 and next steps to take when targeting the specific embryonic day with Cnot1. The project's impact lies in advancing our understanding of CDH pathogenesis, potentially paving the way for targeted therapies aimed at correcting genetic abnormalities implicated in this life-threatening condition.

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