title: Temperature-controlled encapsulation and release of an active enzyme in the cavity of a self-assembled DNA nanocage.
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.
authors.affiliation: Department of Biomedical Engineering, Duke University , Durham, North Carolina 27708, United States.
authors.forename: Sissel
authors.initials: S
authors.lastname: Juul
authors.forename: Federico
authors.initials: F
authors.lastname: Iacovelli
authors.forename: Mattia
authors.initials: M
authors.lastname: Falconi
authors.forename: Sofie L
authors.initials: SL
authors.lastname: Kragh
authors.forename: Brian
authors.initials: B
authors.lastname: Christensen
authors.forename: Rikke
authors.initials: R
authors.lastname: Frøhlich
authors.forename: Oskar
authors.initials: O
authors.lastname: Franch
authors.forename: Emil L
authors.initials: EL
authors.lastname: Kristoffersen
authors.forename: Magnus
authors.initials: M
authors.lastname: Stougaard
authors.forename: Kam W
authors.initials: KW
authors.lastname: Leong
authors.forename: Yi-Ping
authors.initials: YP
authors.lastname: Ho
authors.forename: Esben S
authors.initials: ES
authors.lastname: Sørensen
authors.forename: Victoria
authors.initials: V
authors.lastname: Birkedal
authors.forename: Alessandro
authors.initials: A
authors.lastname: Desideri
authors.forename: Birgitta R
authors.initials: BR
authors.lastname: Knudsen
mesh_terms: D001483:Base Sequence; D002384:Catalysis; D004247:DNA / Q000737:chemistry*; D016503:Drug Delivery Systems; D006735:Horseradish Peroxidase / Q000737:chemistry*; D013058:Mass Spectrometry; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D015394:Molecular Structure; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods; D009690:Nucleic Acid Conformation; D009711:Nucleotides / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013499:Surface Properties; D013696:Temperature; D014357:Trypsin / Q000737:chemistry
chemical_list: D009711:Nucleotides; D009841:Oligonucleotides; D010455:Peptides; D004247:DNA; D006735:Horseradish Peroxidase; D014357:Trypsin