Journal
ACS SENSORS
Volume 7, Issue 1, Pages 60-70Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.1c01579
Keywords
protein-DNA interactions; magnetic modulation biosensing; transcription factor; specificity protein 1; Buttonhead; biosensors; electrophoretic mobility shift assay
Funding
- National Foundation for Practical and Engineering Science [3-14628]
- United States-Israel Binational Science Foundation (BSF) [2009428]
Ask authors/readers for more resources
Identifying and investigating protein-DNA interactions is crucial for biological research. The magnetic modulation biosensing (MMB) system offers a simple and efficient alternative method for detecting these interactions, with faster turnaround times and more accurate results compared to current techniques.
Identifying and investigating protein-DNA interactions, which play significant roles in many biological processes, is essential for basic and clinical research. Current techniques for identification of protein-DNA interactions are laborious, time-consuming, and suffer from nonspecific binding and limited sensitivity. To overcome these challenges and assess protein-DNA interactions, we use a magnetic modulation biosensing (MMB) system. In MMB, one of the interacting elements (protein or DNA) is immobilized to magnetic beads, and the other is coupled to a fluorescent molecule. Thus, the link between the magnetic bead and the fluorescent molecule is established only when binding occurs, enabling detection of the protein-DNA interaction. Using magnetic forces, the beads are concentrated and manipulated in a periodic motion in and out of a laser beam, producing a detectable oscillating signal. Using MMB, we detected protein-DNA interactions between short GC-rich DNA sequences and both a purified specificity protein 1 (Spl) and an overexpressed Buttonhead (BTD) protein in a cell lysate. The specificity of the interactions was assessed using mutated DNA sequences and competition experiments. The assays were experimentally compared with commonly used electrophoretic mobility shift assay, which takes approximately 4-72 h. In comparison, the MMB-based assay's turnaround time is similar to 2 h, and it provides unambiguous results and quantitative measures of performance. The MMB system uses simple and cheap components, making it an attractive alternative method over current costly and time-consuming techniques for analyzing protein-DNA interactions. Therefore, we anticipate that the MMB-based technique will significantly advance the detection of protein-DNA interactions in biomedical research.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available