Multiplexed Mechanotyping of Designer Bonds

Through a complex network of biochemical interactions, transcription factors (TFs) play a crucial role in regulating gene expression. It is known that TFs are able to interact with DNA as well as other proteins in order to recognize DNA sequences and assemble transcription machinery. But as of now, we do not yet have a detailed description of their mechanism of action. Very recently, in 2022, it has been shown that a human transcription factor (KLF4) binds to DNA via a wetting mechanism. We investigate wether similar behaviour is displayed by plant transcription factors. If this is the case, we intend to investigate the kinetics of droplet formation in greater detail. Our approach will be to first develop microfluidics workflows accompanied by fluorescence microscopy to linearize DNA and allow TF binding to be observed. Image analysis tools are then used to generate intensity histograms representing the distribution of TFs along the DNA. Kinetics information is then obtained by analyzing how distributions change in time.