Quantification of transcription factor binding in cell extracts using an electrochemical, structure-switching biosensor.
Transcription factor expression levels, which sensitively reflect cellular development and disease state, are typically monitored via cumbersome, reagent-intensive assays that require relatively large quantities of cells. Here, we demonstrate a simple, quantitative approach to their detection based on a simple, electrochemical sensing platform. This sensor sensitively and quantitatively detects its target transcription factor in complex media (e.g., 250 μg/mL crude nuclear extracts) in a convenient, low-reagent process requiring only 10 μL of sample. Our approach thus appears a promising means of monitoring transcription factor levels.
Department of Chemistry & Biochemistry, University of California, Santa Barbara, California 93106, USA.
Andrew J
AJ
Bonham
Kuangwen
K
Hsieh
B Scott
BS
Ferguson
Alexis
A
Vallée-Bélisle
Francesco
F
Ricci
H Tom
HT
Soh
Kevin W
KW
Plaxco
D015374:Biosensing Techniques / Q000379:methods*; D002457:Cell Extracts / Q000737:chemistry*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D055664:Electrochemical Techniques / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D009690:Nucleic Acid Conformation; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity; D014157:Transcription Factors / Q000032:analysis* / Q000378:metabolism
D002457:Cell Extracts; D014157:Transcription Factors; D004247:DNA