Inhibition of the in vitro replication of DNA by an aptamer-protein complex in an autonomous DNA machine.
DNA replication plays a central role in living organisms. Unregulated or uncontrollable DNA replication is well known to result in many pathological states, such as cancer, autoimmune diseases, and viral/bacterial infections. We report that an aptamer-protein complex could indirectly inhibit in vitro replication of DNA. An isothermal DNA machine based on the strand-displacement amplification is employed to support our assumption. An antithrombin aptamer sequence is rationally encoded into the DNA replication template. Once thrombin binds to the template, the as-formed aptamer-protein complexes can, in turn, become a barrier to the polymerase and inhibit the DNA replication activities in both static and dynamic modes. The inhibition is successfully confirmed by both fluorescence and gel-electrophoresis experiments. Considering the availability of a broad library of aptamers and the existence of various DNA/protein interactions, our results imply the possibility for the rational regulation of DNA replication in vivo.
Zhu
Changfeng
C
Laboratory of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China.
Wen
Yanqin
Y
Li
Di
D
Wang
Lihua
L
Song
Shiping
S
Fan
Chunhai
C
Willner
Itamar
I
D000818:Animals; D052157:Aptamers, Nucleotide / Q000378:metabolism*; D001483:Base Sequence; D002417:Cattle; D004247:DNA / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication; D006801:Humans; D011506:Proteins / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
D052157:Aptamers, Nucleotide; D011506:Proteins; D004247:DNA