Browsing by Subject "c. elegans"
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Item The discovery of novel proteins involved in cell polarity and the PAR complex in c. elegans(2021) Koh, Alexander; Dickinson, DanielCell polarity is a phenomenon that occurs within nearly all organisms. Characterized by the localization of different proteins to different ends of a cell, this process is extremely important for proper biological development. Although studying this mutually antagonistic binding is a growing field of study, cell polarity as a process is still immensely complex and how exactly it is established is still not well known. However, by focusing on known proteins involved in the polarity process, mainly the PAR protein system, other proteins that interact with the PAR complex can be studied for individual effects on cell polarity. The PAR complex was first discovered in c. elegans Multiple experiments were conducted on multiple polarity proteins to identify new potential binding partners to the proteins in the PAR complex. These experiments stem from the cell polarity proteins in the PAR complex, which include aPKC, PAR-6, and PAR-3. A list of proteins from that interact with aPKC was compiled using mass spectrometry pull down. These proteins were studied using RNAi to knock out the gene in C. elegans, and the effects on cell polarity in developing embryos were observed. These genes were then genetically engineered to be injected into worms using Gibson assembly and CRISPR-Cas9 to look for further effects. Furthermore, tests were conducted on strains involving PAR-6. Results show that perhaps the nematode database does not account for an extra intron portray protein sequence, which could lead to further development on the functions of PAR-6, and thus, cell polarity. Finally, mass spectrometry pull down was done on PAR-3 monomer strains due to previous research showing potential of these PAR-3 monomers in having an essential function in C. elegans. This resulted in a list of pulled-down novel proteins, ultimately in which some could potentially affect polarity in new and unknown ways.