Identification and Characterization of Liver-Specific Enhancer Elements in Chromosome 18 of the Nasalis larvatus Genome

Lim, Whye Kit Leonard (2019) Identification and Characterization of Liver-Specific Enhancer Elements in Chromosome 18 of the Nasalis larvatus Genome. Masters thesis, Universiti Malaysia Sarawak.

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Abstract

Enhancers play major roles in the regulation of gene expression at various developmental stages and biological processes temporally and spatially. Albeit the first enhancer was discovered for more than four decades ago, the identification of enhancers is still at its infancy as there is no single universal enhancer mark revealed to date. With the booming emergence of genome sequences of various non-human primates, the study of enhancer landscape across these primates had been made possible recently. Therefore, this study aimed to identify, isolate and characterize enhancers from the genome of one of the most ancient colobine primate, the proboscis monkey (Nasalis larvatus). A total of 13 liver-specific enhancers were identified using three different in silico approaches and they were inserted into expression plasmids before tested further for their activities in HepG2 and A549 cell lines. Two of the strong enhancers and one weak enhancer were subjected to deletion analysis and activity determination subsequently in both cell lines. This study has discovered several clusters of adjacently located transcription factor binding sites that influence enhancer activities and tissue specificities greatly. The results from this research will drive future investigations on the enhancer landscape of the proboscis monkey and comparison of the landscape across different primates. Keywords: Enhancer, identification, proboscis monkey, in silico, expression

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