FUNCTIONAL CHARACTERIZATION OF TRANSCRIPTION FACTORS IN DEVELOPMENTAL REGULATION

Abstract

Transcription factors (TFs) have a significant role in regulating gene expression in the developmental processes. Exact control of these variables is critical to the correct differentiation of cells, the formation of tissues and the development of organisms. Nevertheless, the traditional research usually does not provide dynamic and quantitative descriptions of the TF activity in different regulatory conditions. This paper is a functional characterization of key developmental regulation transcription factors in engineered regulatory constructs in mammalian cells. HEK293 cells were introduced to transient transfection methods to incorporate TF-regulated systems of expression. Reporter assays (GFP and luciferase) were used to monitor gene expression as well as RT-qPCR and Western blotting of protein levels. Activation of transcription factors with dose dependence, time dynamics, and stability through regulation were assessed. The findings have shown that the regulatory behavior of transcription factors is highly tunable and condition-dependent, and is more precise and less variable in controlled conditions. These results suggest the possibilities of transcription factor-based regulatory systems in development biology, regenerative medicine, and therapeutic gene regulation.