Presentation description

ZBTB4 is a methyl-CpG binding protein that regulates epigenetic-based transcriptional processes and is comprised of two separated zinc finger (ZF) domains involved in nucleic acid binding. ZBTB4 acts as an epigenetic reader through methylated DNA recognition by the N-terminal (N-term) ZFs, however, a subset of identified ZBTB4 genomic targets contain elements correlated to potential quadruplex forming sequences (PQS), and do not correspond to N-term ZF domain readout. Importantly, our lab has discovered that ZBTB4 can also bind DNA G-quadruplex (G4) motifs through its C-terminal (C-term) ZF domain, thus suggesting a role for ZBTB4 in genomic G4-mediated transcriptional regulation. Genomic G4s can affect cellular processes and they have been correlated with disease states like cancer, which highlights ZBTB4's potential role in modulating gene expression in response to genomic G4s. In the Buck lab we aim to characterize this ZBTB4:G4 interaction. To advance this research, my efforts focused on implementing a new method to enzymatically produce G4-producing single-stranded DNA (ssDNA) oligonucleotides that can be utilized for downstream ZBTB4 binding experiments. Although ssDNA oligonucleotides are commonly produced through chemical synthesis, enzymatic production can produce a higher yield and quality of oligonucleotides in less time. In addition, molecular cloning of different ZBTB4 C-term construct was implemented. Together, these efforts will provide methodologies for producing G4 DNAs and protein constructs, further aiding the investigation of ZBTB4's role in G4-related transcriptional regulation.