Synthetic nanostructures including nucleic acids and/or other entities

Abstract

Articles, compositions, kits, and methods relating to nanostructures, including synthetic nanostructures, are provided. Certain embodiments described herein include structures having a core-shell type arrangement; for instance, a nanostructure core may be surrounded by a shell including a material, such as a lipid bilayer, and may include other components such as oligonucleotides. In some embodiments, the structures, when introduced into a subject, can be used to deliver nucleic acids and/or can regulate gene expression. Accordingly, the structures described herein may be used to diagnose, prevent, treat or manage certain diseases or bodily conditions. In some cases, the structures are both a therapeutic agent and a diagnostic agent.

1 .- 5 . (canceled) 
     
     
         6 . A method for regulating gene expression comprising:
 delivering a nanostructure to a subject or a biological sample in an effective amount for regulating gene expression in the subject or biological sample,
 wherein the nanostructure comprises:
 a core; 
 a shell comprising a lipid surrounding the core or a hydrophobic shell surrounding the core; and 
 
 an oligonucleotide adapted to regulate gene expression associated with at least a portion of the shell, 
 wherein the nanostructure is adapted to sequester cholesterol. 
   
     
     
         7 .- 99 . (canceled) 
     
     
         100 . The method of  claim 6 , wherein the oligonucleotide is adsorbed to a surface of the shell. 
     
     
         101 . The method of  claim 6 , wherein the oligonucleotide is physisorbed onto a portion of the shell. 
     
     
         102 . The method of  claim 6 , wherein the oligonucleotide is chemisorbed onto a portion of the shell, 
     
     
         103 . The method of  claim 6 , wherein the oligonucleotide is attached to a portion of the shell through ionic, hydrophobic, hydrophilic, electrostatic, or van der Waals interactions. 
     
     
         104 . The method of  claim 6 , wherein the oligonucleotide is attached to a surface of the core through an intervening layer such as a passivating layer. 
     
     
         105 . The method of  claim 6 , wherein the oligonucleotide is adsorbed to an inner portion, outer portion, interior portion of the shell, and/or combinations thereof. 
     
     
         106 . The method of  claim 6 , wherein the oligonucleotide is covalently or near-covalently bonded to the core or to the shell. 
     
     
         107 . The method of  claim 6 , wherein the oligonucleotide is an end-modified oligonucleotide. 
     
     
         108 . The method of  claim 105 , wherein the oligonucleotide is a thiol end-modified oligonucleotide, wherein the oligonucleotide is attached to the core through a thiol-metal bond. 
     
     
         109 . The method of  claim 105 , wherein the oligonucleotide is a cholesterol-modified oligonucleotide. 
     
     
         110 . The method of  claim 6 , wherein the oligonucleotide is single-stranded. 
     
     
         111 . The method of  claim 6 , wherein the oligonucleotide is double-stranded. 
     
     
         112 . The method of  claim 6 , wherein the oligonucleotide comprises antisense DNA, siRNA, or microRNA. 
     
     
         113 . The method of  claim 6 , wherein the oligonucleotide comprises mRNA. 
     
     
         114 . The method of  claim 6 , wherein the oligonucleotide has a length of about 8 to about 500 nucleotides or base pairs in length. 
     
     
         115 . The method of  claim 6 , wherein the shell comprises a lipid bilayer. 
     
     
         116 . The method of  claim 115 , wherein the lipid bilayer comprises 50-200 phospholipids. 
     
     
         117 . The method of  claim 6 , wherein at least a portion of the lipid bilayer is physisorbed to the core. 
     
     
         118 . The method of  claim 6 , wherein the nanostructure further comprises a protein associated with at least a portion of the structure. 
     
     
         119 . The method of  claim 118 , wherein the structure comprises an apolipoprotein surrounding the core. 
     
     
         120 . The method of  claim 6 , wherein the structure has low endosomal sequestration defined by at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% of nanostructures in a cell being located within the cytoplasm or nucleus of the cell. 
     
     
         121 . The method of  claim 6 , wherein the nanostructure has a binding constant to cholesterol, K d , of about 10 mM or less. 
     
     
         122 . The method of  claim 6 , wherein the structure has a largest cross-sectional dimension of less than or equal to about 50 nm, 35 nm, or 30 nm. 
     
     
         123 . The method of  claim 6 , wherein the structure has a zeta potential of less than or equal to −10 mV, −20 mV, or −30 mV. 
     
     
         124 . The method of  claim 123 , wherein the structure has a zeta potential of less than or equal to −30 mV. 
     
     
         125 . The method of  claim 6 , wherein the structure is administered in a single or divided dose according to a dosing schedule. 
     
     
         126 . The method of  claim 6 , wherein the structure is administered in two or more doses. 
     
     
         127 . The method of  claim 6 , wherein the subject has cancer or a cell in the biological sample is associated with a cancer. 
     
     
         128 . The method of  claim 127 , wherein the cancer is prostate cancer. 
     
     
         129 . A method for promoting cellular uptake of an oligonucleotide, the method comprising:
 delivering a nanostructure to a subject or a biological sample in an effective amount for promoting cellular uptake of the oligonucleotide in the subject or biological sample,   wherein the nanostructure comprises:
 a core; 
 a shell comprising a lipid surrounding the core or a hydrophobic shell surrounding the core; and 
   an oligonucleotide adapted to regulate gene expression associated with at least a portion of the shell,   wherein cellular uptake of the oligonucleotide results in regulation of gene expression in the cell,   wherein the nanostructure is adapted to sequester cholesterol.   
     
     
         130 . The method of  claim 129 , wherein the nano structure comprises a plurality of oligonucleotides adapted to regulate gene expression, wherein the oligonucleotides have greater than 80%, 85%, 90%, 95%, 97%, or 99% cellular uptake.