Abstract

There is a vast diversity in tetrapod lung branching morphology. Phylogenetically, much of the pulmonary diversity among vertebrates appears to arise from the way epithelial tubes branch or form saccular (cyst) structures. Fgf10 activity has been shown to play a critical role in regulating branch versus cyst morphology. We hypothesize that the species-specific differences in lung morphology may be primarily due to species-specific differences in Fgf10 expression. To test this hypothesis, we have performed bioinformatic analyses on the Fgf10 locus and have identified a conserved 11 kb noncoding region that potentially contains the Fgf10 lung enhancer. We are taking a large DNA sequence upstream of the Fgf10 gene of the American Alligator and swapping it into the orthologous locus in the genome of chicken primordial germ cells (cPGCs). We are accomplishing these swaps by using a combination of homology directed repair (HDR) and recombinase mediated cassette exchange (RMCE) in cPGCs. These edited cell lines can be used to generate germline chimeric chickens capable of producing offspring that putatively drive Fgf10 expression in the lung under control of regulatory sequences from various other reptiles. We have also generated a cPGC line where, through RMCE, we can easily target any enhancer from any organism to drive a GFP reporter as a means to test the temporal and spatial regulatory characteristics of these enhancers. This work is funded through a BYU Turkey Vaccine Grant and a Skaggs Mentoring Grant.

Degree

MS

College and Department

Life Sciences

Rights

https://lib.byu.edu/about/copyright/

Date Submitted

2022-11-22

Document Type

Thesis

Handle

http://hdl.lib.byu.edu/1877/etd12605

Keywords

Fgf10, evolution, transgenic, chicken, alligator, lung, RMCE, HDR

Language

english

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