Polymer nanoparticle and dna nanostructure compositions and methods for non-viral delivery

Abstract

The invention relates to polymer nanoparticle and DNA nanostructure delivery compositions for non-viral delivery, and methods therefor. More particularly, the invention relates to polymer nanoparticle delivery compositions, such as reversible addition-fragmentation chain transfer (RAFT) polymer compositions, and DNA nanostructure delivery compositions, such as DNA origami compositions, for the delivery of more than one payload, or for the delivery of a nucleic acid construct payload of 3 kB or more, and methods therefor.

What is claimed is: 
     
         1 . A DNA nanostructure delivery composition comprising: i) a single stranded DNA scaffold and ii) one or more oligonucleotides that bind through complementary base pairing with a segment of the DNA scaffold, wherein the one or more oligonucleotides cause the DNA scaffold to fold, and wherein the composition comprises more than one payload for delivery or a nucleic acid construct payload of 3 kB or more. 
     
     
         2 . The composition of  claim 1  wherein the more than one payload comprises nucleic acids. 
     
     
         3 . The composition of  claim 2  wherein the nucleic acids comprise DNA or RNA. 
     
     
         4 . The composition of  claim 2  wherein the nucleic acids comprise a ribonucleoprotein. 
     
     
         5 . The composition of  claim 2  wherein the payload nucleic acids are used for homology directed repair or as transposable elements. 
     
     
         6 . The composition of  claim 2  wherein the payload nucleic acids comprise a short guide RNA (sgRNA) and a donor DNA strand. 
     
     
         7 . The composition of  claim 6  wherein the sgRNA is used for targeting an enzyme to a specific genomic sequence. 
     
     
         8 . The composition of  claim 1  wherein the payloads comprise a CRISPR associated enzyme. 
     
     
         9 . The composition of  claim 7  wherein the targeted enzyme is a CRISPR associated enzyme. 
     
     
         10 . The composition of  claim 1  wherein the payloads comprise a CRISPR associated enzyme, an sgRNA, and a donor DNA strand. 
     
     
         11 . The composition of  claim 1  wherein the payloads comprise CRISPR/Cas9. 
     
     
         12 . The composition of  claim 1  wherein the payloads comprise CRISPR/Cas9, an sgRNA, and a donor DNA strand. 
     
     
         13 . The composition of  claim 1  wherein the payload comprises CRISPR/Cas9 and Cas9 is fused with a deaminase. 
     
     
         14 . The composition of  claim 1  wherein the payloads comprise a coding sequence for Cas9, an sgRNA, and a donor DNA strand in the form of a plasmid. 
     
     
         15 . The composition of  claim 1  wherein the payloads consist of one molecule each of CRISPR/Cas9, an sgRNA, and a donor DNA strand. 
     
     
         16 . The composition of  claim 1  wherein the nucleic acid construct payload of 3 kB or more comprises a CAR-T DNA construct. 
     
     
         17 . The composition of  claim 1  wherein the payloads comprise an antisense oligonucleotide. 
     
     
         18 . The composition of  claim 1  wherein the nucleic acid construct payload is of a size selected from the group consisting of 3 kB or more, 3.5 kB or more, 4 kB or more, 4.5 kB or more, 5 kB or more, 5.5 kB or more, 6 kB or more, 6.5 kB or more, 7 kB or more, 7.5 kB or more, 8 kB or more, and 8.5 kB or more. 
     
     
         19 . The composition of  claim 1  wherein the payloads comprise a reverse transcriptase. 
     
     
         20 . The composition of  claim 1  wherein the DNA scaffold and the one or more oligonucleotides comprise M13 bacteriophage DNA.