| dc.description.abstract | The pursuit of effective tools for probing biological systems has led to a growing
interest in approaches that offer both rapid, reversible, and accurate methods.
Specifically, the ability to perturb the function of specific proteins with precision is
highly sought after, as it contributes significantly to our capacity to unravel the intricacies
of complex biological processes. Effective methods to regulate gene expression or protein
function within mammalian hosts are essential for understanding basic biological
mechanisms (Banaszynski 2008). Examples of these methods include generating animal
knockouts, siRNA, the Cre-loxP system, and small molecule activators or inhibitors.
However, these methods are limited because they may not be reversible, are non-specific,
or—in the case of animal knockouts—the gene being knockout out is essential for
development thus confounding the study of the protein’s function. Another method
involves synthetic transcription factors paired with destabilizing-domain fusion proteins
that target a gene of interest. The destabilizing-domain are stabilized in the presence of a
ligand, thus providing the ability to control and regulate gene expression. This technique
can provide the regulatability, specificity, and ease of use in understanding complex
biological mechanisms. One component of synthetic transcription factors is the design of
activation domains utilized. Activation domains promote recruitment of chromatin
modifiers, which cause chromatin decondensation and accumulation of histone marks,
thus promoting transcription (Martella 2021). The variability of transcription efficacy is
based on the properties of different activation domains. In this study, three different
domains are investigated: VPR, VP64, and p65, with the goal of determining which one
provides the highest dynamic range in a lentiviral-transduced mammalian cell system
using a synthetic transcription factor fused to a drug-dependent destabilizing protein
domain. | |
