A zinc finger protein array for the visual detection of specific DNA sequences for diagnostic applications.
The visual detection of specific double-stranded DNA sequences possesses great potential for the development of diagnostics. Zinc finger domains provide a powerful scaffold for creating custom DNA-binding proteins that recognize specific DNA sequences. We previously demonstrated sequence-enabled reassembly of TEM-1 β-lactamase (SEER-LAC), a system consisting of two inactive fragments of β-lactamase each linked to engineered zinc finger proteins (ZFPs). Here the SEER-LAC system was applied to develop ZFP arrays that function as simple devices to identify bacterial double-stranded DNA sequences. The ZFP arrays provided a quantitative assay with a detection limit of 50 fmol of target DNA. The method could distinguish target DNA from non-target DNA within 5 min. The ZFP arrays provided sufficient sensitivity and high specificity to recognize specific DNA sequences. These results suggest that ZFP arrays have the potential to be developed into a simple and rapid point-of-care (POC) diagnostic for the multiplexed detection of pathogens.
Genome Center, Department of Pharmacology, University of California, Davis, CA 95616, USA.
Moon-Soo
MS
Kim
Gulnaz
G
Stybayeva
Ji Youn
JY
Lee
Alexander
A
Revzin
David J
DJ
Segal
D004269:DNA, Bacterial / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D025202:Molecular Diagnostic Techniques; D040081:Protein Array Analysis / Q000379:methods*; D015202:Protein Engineering; D017422:Sequence Analysis, DNA; D016335:Zinc Fingers; D001618:beta-Lactamases / Q000235:genetics
D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D001618:beta-Lactamases; C034413:beta-lactamase TEM-1