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ORCID
https://orcid.org/0000-0002-9523-1463
Access Type
Open Access Thesis
Document Type
thesis
Degree Program
Molecular & Cellular Biology
Degree Type
Master of Science (M.S.)
Year Degree Awarded
2019
Month Degree Awarded
September
Abstract
LBH is a highly conserved protein whose role during vertebrate development is relatively under-studied. In collaboration with the Albertson lab, our lab has previously shown that it is necessary for cranial neural crest cell migration in the zebrafish and in Xenopus laevis. The molecular mechanisms through which it acts are not well understood.
In Xenopus, LBH is a maternally deposited protein. As such, studying its role in early development has not been feasible through the morpholino-mediated knockdown techniques that prevent translation of target genes. Recently, a technique for degrading endogenous proteins was developed, called Trim-Away. This was developed in mammalian systems and utilizes the E3 ubiquitin ligase Trim21 in conjunction with an antibody against a protein of interest in order to degrade the protein. In order to observe the effects of a knockdown of LBH during early embryonic development, we sought to modify the technique for use in Xenopus. We injected embryos with mRNA encoding the human form of trim21 along with a monoclonal antibody against LBH that our lab developed (2B8) and tracked degradation of the protein over time, monitoring embryos for any phenotypes arising during early development.
Our results demonstrate that Trim-Away can be utilized in Xenopus. LBH depleted embryos display a variety of defects during gastrulation, the process by which the three germ layers are properly organized. These appear to be mainly due to defects in fibronectin fibrillogenesis and mesodermal migration.
DOI
https://doi.org/10.7275/15206577
First Advisor
Helene Cousin
Second Advisor
Dominique Alfandari
Third Advisor
R. Craig Albertson
Recommended Citation
Weir, Emma, "Investigating The Role Of LBH During Early Embryonic Development In Xenopus Laevis" (2019). Masters Theses. 860.
https://doi.org/10.7275/15206577
https://scholarworks.umass.edu/masters_theses_2/860