Oligonucleotide-directed self-assembly of proteins: semisynthetic DNA—streptavidin hybrid molecules as connectors for the generation of macroscopic arrays and the construction of supramolecular bioconjugates

Abstract

ABSTRACT

A Protein‐Interaction Array Inside a Living Cell

“…To create a pattern of bait-PARCs inside cells, we used DNA-directed immobilization (DDI) [4] to generate micrometer-scale arrays of antibodies with binding specificity for the peptide epitope on the bait-PARC. The DDI method takes advantage of specific hybridization of complementary oligonucleotides and thereby allows the sitespecific capture of sensitive biomolecules by the DNA microstructures on a solid substrate under mild conditions.…”

Peptide-based biopolymers in biomedicine and biotechnology

“…In addition, bionanofabrication potentially allows direct interrogation of numerous biomolecules in a parallel manner. In materials science, molecular recognition of biomolecules such as DNA and antibodies can direct the assembly of nanoscale components such as gold nanoparticles, quantum dots, and proteins [23,[343][344][345][346]. Generation of more complex biomolecular ensembles is also possible when using self-assembling biomolecules.…”

A polycatenated DNA scaffold for the one-step assembly of hierarchical nanostructures

“…Two micromolar 1 was reacted with 2 M 2 in ligation buffer consisting of 66 mM Tris⅐HCl, 10 mM MgCl 2, 1 mM ATP, 15% polyethylene glycol (PEG 6,000), 1 mM DTT, pH 7.6. The mixture was first heated to 90°C for 10 min then cooled to 50°C for different time intervals (30,60, and 90 min, respectively), fast cooled to 25°C, and exposed to ligase 200 units/l for 30 min. The resulting mixture was loaded in 0.6% agarose gel and exposed with SYBR Green I.…”

“…DNA, however, is a robust biopolymer that, through Watson-Crick base pairing and the multitude of sequences that can be formulated, might yield self-assembled protein scaffolds. Indeed, DNA has been used for the organization of proteins (9,10) and elegant, topologically complex 3D architectures, such as Borromean rings (11), nanotubes (12), [2]catenanes (13), 2D tilings (14)(15)(16)(17)(18), and chains (19), onto which nanoparticles (20)(21)(22)(23)(24)(25)(26)(27)(28) and proteins (19,20,(29)(30)(31)(32) have been tethered by using complementary single-stranded DNA (ssDNA), biotin-streptavidin interactions, or gold-thiol bonds. These studies, however, have been limited to the immobilization of a single biomolecule or nanoparticle onto the scaffold or require multistep protocols to organize more than one nanomaterial onto the template.…”