Use of DNA origami nanostructures for molecular information based data storage systems

A library comprising a plurality of origami folded DNA data storage files (DNAFiles), each of said DNAFiles comprising

• a single stranded DNA scaffold;
• and
• a plurality of single stranded DNA staple oligonucleotides that each bind through complementary base pairing with two non-contiguous nucleic acid sequences of the DNA scaffold, wherein said staple oligonucleotides cause the DNA scaffold to fold into a two or three dimensional shape having a first surface;
• a plurality of data oligonucleotides, said data oligonucleotides comprising a sequence complementary to a nucleic acid sequence of said single stranded DNA scaffold, a nucleic acid sequence that encodes digital information, and a first primer binding sequence and a second primer binding sequence, wherein the first primer binding sequence is 5′ to the digital information encoding nucleic acid sequence, the second primer binding sequence is 3′ to the digital information encoding nucleic acid sequence, wherein said plurality of data oligonucleotides are localized to said first surface, wherein the individual DNAFiles differ from one another based on the nucleic acid sequences of the plurality of data oligonucleotides bound to the DNA scaffold of each DNAFile wherein the data oligonucleotides of each individual origami folded DNAFile of said library further comprise an identical set of PCR binding sequences for preselected PCR primers, where the PCR binding sequences differ between the data oligonucleotides of each respective DNAFile of the library.

A method of storing digital information using DNA as the storage medium, said method comprising the steps:

• providing a single stranded DNA scaffold;
• and
• providing a plurality of single stranded staple oligonucleotides that each bind through complementary base pairing with two non-contiguous nucleic acid sequences of the DNA scaffold, wherein said staple oligonucleotides cause the DNA scaffold to fold into a two or three dimensional shape having a plurality of external surfaces;
• mixing said DNA scaffold and said staple oligonucleotides under conditions that allow sequence specific hybridization of the staple oligonucleotides to the DNA scaffold and folding of the DNA scaffold;
• and
• hybridizing a plurality of data oligonucleotides exclusively to one external surface of said plurality of external surfaces to store digital information using said data oligonucletides as the storage medium, wherein said data oligonucleotides comprise a sequence complementary to a segment of said single stranded DNA scaffold, a nucleic acid sequence that encodes digital information, a first primer binding sequence and a second primer binding sequence, wherein the first primer binding sequence is locate 5′ to the digital information encoding nucleic acid sequence, and the second primer binding sequence is locate 3′ to the digital information encoding nucleic acid sequence.

A library comprising a plurality of origami folded DNA data storage files (DNAFiles), each of said DNAFiles comprising

• a single stranded DNA scaffold;
• and
• a plurality of single stranded DNA staple oligonucleotides that each bind through complementary base pairing with two non-contiguous nucleic acid sequences of the DNA scaffold, wherein said staple oligonucleotides cause the DNA scaffold to fold into the shape of a multi-layered sheet having a top surface and a bottom surface;
• a plurality of data oligonucleotides, said data oligonucleotides comprising a sequence complementary to a nucleic acid sequence of said single stranded DNA scaffold, a nucleic acid sequence that encodes digital information, and a first primer binding sequence and a second primer binding sequence, wherein the first primer binding sequence is 5′ to the digital information encoding nucleic acid sequence, the second primer binding sequence is 3′ to the digital information encoding nucleic acid sequence, wherein said plurality of data oligonucleotides are bound only to the top surface, further wherein the individual DNAFiles differ from one another based on the nucleic acid sequences of the data oligonucleotides bound to the DNA scaffold of each DNAFile.