Topogami and method for making interlocked single stranded dna rings
AssigneeUniwersytet Jagiellonski
InventorsJonathan HEDDLE, Yusuke SAKAI, Gerrit WILKENS
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Abstract
It has been disclosed a method for the construction of single stranded DNA catenanes at the kilobase length scale, which can be used for the assembling scaffolds of covalently connected DNA origamis.
Claims15
A nucleic acid nanostructure (topogami) characterised in that it is artificial nucleic acid nanostructure consisting of at least 2 interlocked single stranded polynucleotide rings and associated complementary staples strands capable of folding into arbitrary 2D or 3D structures, wherein each interlocked single stranded polynucleotide ring consists of more than 300 nucleotides.
A method for making interlocked single stranded DNA rings whereby double stranded rings comprising following steps:
- a) engineering of DNA plasmid vectors to produce a vector consisting of two scaffold domains connected by parallel recognition sequences (res) in a negatively supercoiled parent plasmid and having integrated on opposite strands of the vector and having two nickase sites, one each on opposite strands of the vector;
- b) processing of the plasmid obtained in step a) by Tn3 resolvase, a serine recombinase to catalyse a site-specific recombination at the res sites to form a supercoiled catenane;
- c) conversion of catenated product obtained in step b) into ssDNA by Nt.BspQI nicking and subsequent exonuclease digestion, with nicked non-catenated parent plasmid removed by exonuclease digestion;
- d) purification of the catenated ssDNA product by phenol:chloroform extraction and concentration by EtOH precipitation.
Dependencies are inferred from the claim text.