Investigation of the Role of LSD1 Variants in Complete Embryonic Stem Cell Differentiation and Cellular Function
Marino, Maxim E.
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The human body compresses DNA by wrapping it around a set of proteins known as histones, forming a complex known as a nucleosome. To compensate for the reduction in DNA availability, numerous biological processes exist that allow for this DNA to temporarily become more accessible to other proteins such as transcription factors. This is possible through the utilization of histone-modifying proteins. One such protein is lysine-specific demethylase 1 (LSD1), which catalyzes the demethylation of lysine 4 on histone 3 for a given nucleosome. It is thought that LSD1 may play a critical role in the complete differentiation of embryonic stem cells through the suppression of enhancer DNA sequences, and that genetic mutations in LSD1 may be the cause of a currently unnamed neurodevelopmental disorder. The ability of LSD1 variants to rescue cellular function in LSD1-knockout embryonic stem cells was investigated in order to obtain information regarding its mechanism of action. This was to be accomplished by comparing qPCR analysis of wild type and LSD1-knockout mouse embryonic stem cell cultures with knockout cultures containing a reintroduction of LSD1. Error in experimental design caused an incomplete knockout of LSD1 in all knockout cultures, and qualitative data suggests that none of the cultures had significant function rescued. The mechanism of action of LSD1 will be further investigated through an overexpression system comparing the wild type and catalytically dead versions of LSD1.