MURINE TRANSGENE INSERTIONAL MUTATION INVOLVING RUNX1T1 AND GM11823 GENES AND THEIR CONTRIBUTION TO CLEFT PALATE AND RIB ANOMALIES Public Deposited

Downloadable Content

Download PDF
Last Modified
  • June 7, 2019
Creator
  • Alazzam, Melanie
    • Affiliation: School of Dentistry, Oral and Craniofacial Biomedicine PhD Program
Abstract
  • Objective: Cleft Palate (CP) is a common birth defect in humans occurring in 6.35/10,000 live births and it has been repeatedly shown that animal models are useful in dissecting molecular etiologies of CP. The OVE1328 mouse line develops CP as a consequence of transgene (Tg) insertion mutagenesis. Preliminary data shows Tg complex integration at chromosome 4 band A2. The goal of this work was to characterize the mutation in the OVE1328 transgenic mouse line. Methods: Genotyping microarrays and RNA-seq were used to identify the Tg insertion site. The insertion site was further validated by conventional PCR. Histology and skeletal staining were used to phenotype CP (OVE1328 (Tg/Tg)) and Wt embryos. Results: In OVE1328 embryos the transgene disrupts Runx1t1 gene at intron 12 (13,876,840 bp). The integration is associated with a deletion mutation of part of intron 12 and whole exons 13 and 14 of Runx1t1. Runx1t1 (also known as Cbfa2t1h, Eto, Mtg8) is mainly studied for its role in acute myelogenous leukemia as a fusion gene in humans and gut development in mice. The Tg insertion disruption extends to the Gm11823 gene which encodes a long non coding RNA. Little is known regarding the normal function and expression of Gm11823. The Tg integrates at intron 2 (13,949,713 bp) of Gm11823 and induces the ectopic expression of an altered message. Homozygous disruption of both genes is associated with CP (100% of OVE1328 (Tg/Tg) embryos) and rib anomalies (supernumerary ribs, 86% of OVE1328 (Tg/Tg) embryos. Furthermore, OVE1328 (Tg/Tg) embryos are ~13% smaller by weight than Wt and OVE1328 (Tg/+) littermates indicting growth delay. Supported by NIH/NIDCR DE015180.
Date of publication
Keyword
Subject
Identifier
Resource type
Rights statement
  • In Copyright
Advisor
  • Frazier-Bowers, Sylvia
  • Everett, Eric
  • Van Aalst, John
  • Rossouw, Paul
  • Wright, John
Degree
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2014
Language
Publisher
Place of publication
  • Chapel Hill, NC
Access
  • This item is restricted from public view for 2 years after publication.
Parents:

This work has no parents.

Items