DNase I Footprinting

DNase I Footprinting

Introduction

Deoxyribonuclease I (DNase I) footprinting is a valuable technique for locating the specific binding sites of proteins on DNA. It can not only find the target protein that binds to specific DNA, but also identify the sequence that the target protein binds to. The basis of this assay is that the bound protein can protect the phosphodiester backbone of DNA from DNase I hydrolysis. First, use DNase I to digest the radioactively-labeled or fluorescently-labeled DNA fragments, and then use gel electrophoresis to detect the cleavage pattern. If the protein binds to the DNA fragment, the binding site will be protected from enzymatic cleavage, and a clear area called "footprint" will be formed on the gel.

At present, the footprint method has been developed into a quantitative method that can be used to determine the separate binding curve of individual protein-binding sites on DNA. For sites that interact cooperatively, simultaneous numerical analysis of all binding curves can be used to analyze the intrinsic binding and cooperative components of these energies. Nowadays, although transcriptional regulation has been extensively studied, there are still many unknowns. Transcription factors and related proteins that drive or inhibit transcription together with promoters, enhancers, or silencers are essential for understanding the regulation of individual genes in the genome. Technologies such as DNase I footprint can effectively help us reveal the complexity of transcription control.

DNase I footprinting theoryFigure 1. DNase I footprinting theory (Carey, M.F.; et al. 2013)

Services

Creative Proteomics has launched the DNase I footprinting platform for nearly two decades. During this period, our experienced experts and technicians have built the platform into a high-quality, first-class technical service center through continuous research and upgrade. We have achieved multi-platform cooperation, aiming to help customers complete projects more efficiently. The DNase I footprinting experiment we provide includes but is not limited to the following steps:

DNase-I-Footprinting-Platform-2

  • Pour gel from premade acrylamide/urea mix
  • Prepare buffers for DNase I footprinting
  • DNA-binding reactions
  • DNase I footprinting
  • Gel electrophoresis and autoradiography
  • Data analysis
  • Generate a dose-response curve
  • Dose-response data analysis

Customers can choose different technology platforms according to project requirements, or contact us directly for consultation, and our expert team will provide you with customized experimental procedures.

Creative Proteomics is an international biotechnology company dedicated to research in molecular interactions and other related fields. The DNase I footprinting platform we constructed has the characteristics of high quality and efficiency, and the data obtained can be directly used for paper publication. Our one-stop service aims to save customers time and money.

References

  1. Carey, M.F.; et al. DNase I Footprinting. Cold Spring Harbor Protocols. 2013.
  2. Brenowitz, M.; et al. DNase I Footprint Analysis of Protein-DNA Binding. Current Protocols in Molecular Biology. 1989.
* This service is for RESEARCH USE ONLY, not intended for any clinical use.

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