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Temperature-Controlled Encapsulation and Release of an Active Enzyme in the Cavity of a Self-Assembled DNA Nanocage

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ABSTRACT We demonstrate temperature-controlled encapsulation and release
of the enzyme horseradish peroxidase using a preassembled and covalently closed
three-dimensional DNA cage structure as a controllable encapsulation device. The
utilized cage structure was covalently closed and composed of 12 double-stranded
B-DNA helices that constituted the edges of the structure. The double stranded helices
were interrupted by short single-stranded thymidine linkers constituting the cage corners except for one, which was composed by four 32 nucleotide long
stretches of DNA with a sequence that allowed them to fold into hairpin structures. As demonstrated by gel-electrophoretic and fluorophore-quenching
experiments this design imposed a temperature-controlled conformational transition capability to the structure, which allowed entrance or release of an
enzyme cargo at 37 C while ensuring retainment of the cargo in the central cavity of the cage at 4 C. The entrapped enzyme was catalytically active inside the
DNA cage and was able to convert substrate molecules penetrating the apertures in the DNA lattice that surrounded the central cavity of the cage.
OriginalsprogEngelsk
TidsskriftACS Nano
Vol/bind7
Nummer11
Sider (fra-til)9724-9734
Antal sider11
ISSN1936-0851
DOI
StatusUdgivet - 29 okt. 2013

    Forskningsområder

  • DNA nanocage, controlled encapsulation and release, drug delivery, temperature, MD simulation, fluorescence-quenching, self-assembly

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