Nucleic acid particles, methods and use thereof

Abstract

The present invention provides, among other things, a particle which includes a core comprised of self-assembled one or more nucleic acid molecules, the core being characterized by an ability to adopt at least two configurations: a first configuration having a first greatest dimension greater than 2 μm and; a second configuration having a second greatest dimension less than 500 nm, wherein addition of a film coating converts the core from its first configuration to its second configuration. Methods of making and using of provided particles are also disclosed.

We claim: 
     
         1 . A particle, comprising:
 a core comprised of self-assembled one or more nucleic acid molecules, the core being characterized by an ability to adopt at least two configurations:   a first configuration having a first greatest dimension greater than 2 μm and;   a second configuration having a second greatest dimension less than 500 nm,   wherein addition of a film coating converts the core from its first configuration to its second configuration.   
     
     
         2 . The particle of  claim 1 , wherein the core contains a single nucleic acid molecule. 
     
     
         3 . The particle of  claim 1 , wherein the core is comprised of a plurality of nucleic acid molecules. 
     
     
         4 . The particle of  claim 3 , wherein individual nucleic acid molecules within the core have different nucleic acid sequences. 
     
     
         5 . The particle of  claim 3 , wherein all nucleic acid molecules within the core have substantially the same nucleic acid sequence. 
     
     
         6 . The particle of  claim 3 , wherein nucleic acid molecules within the core have sequences that share at least one common sequence element. 
     
     
         7 . The particle of  claim 1 , wherein at least one nucleic acid molecule within the core has a nucleotide sequence that comprises multiple copies of at least a first sequence element. 
     
     
         8 . The particle of  claim 1 , wherein at least one nucleic acid molecule within the core has a nucleotide sequence that comprises multiple copies of each of at least a first and a second sequence element. 
     
     
         9 . The particle of  claim 8 , wherein the at least one nucleic acid molecule has a nucleotide sequence that comprises alternating copies of the first and second sequence elements. 
     
     
         10 . The particle of  claim 8 , wherein the at least one nucleic acid molecule has a nucleotide sequence that comprises multiple copies of each of three or more sequence elements. 
     
     
         11 . The particle of  claim 1 , wherein at least one nucleic acid molecule has a nucleotide sequence that includes one or more sequence elements found in a natural source. 
     
     
         12 . The particle of  claim 11 , wherein the at least one nucleic acid molecule has a nucleotide sequence that includes a first sequence element that is found in a first natural source and a second sequence element that is found in a second natural source. 
     
     
         13 . The particle of  claim 12 , wherein the first and second natural sources are the same. 
     
     
         14 . The particle of  claim 12 , wherein the first and second natural sources are different. 
     
     
         15 . The particle of  claim 1 , wherein at least one nucleic acid molecule in the core has a nucleotide sequence that represents an assemblage of sequence elements found in one or more source nucleic acid molecules. 
     
     
         16 . The particle of  claim 15 , wherein the at least one nucleic acid molecule has a nucleotide sequence that represents an assemblage of at least two different sequence elements found in two different source nucleic acid molecules. 
     
     
         17 . The particle of  claim 1 , wherein at least a portion of the nucleic acid molecules within a core is cleavable. 
     
     
         18 . The particle of  claim 1 , wherein the nucleic acid molecules within a core comprise single-stranded, double-stranded, triple-stranded nucleic acids or combination thereof. 
     
     
         19 . The particle of  claim 1 , wherein the nucleic acid molecules within a core are arranged in a crystalline structure comprising lamellar sheets. 
     
     
         20 . The particle of  claim 1 , wherein the nucleic acid molecules within a core are formed via amplification by rolling circle amplification (RCA), rolling circle transcription (RCT) or both. 
     
     
         21 . The particle of  claim 1 , wherein the nucleic acid molecules within a core comprise a stem-loop or linear structure. 
     
     
         22 . The particle of  claim 1 , wherein the core comprises about 1×10 3  to 1×10 8  copies of a sequence element. 
     
     
         23 . The particle of  claim 1 , wherein the core comprises at least 1×10 6  copies of a sequence element. 
     
     
         24 . The particle of  claim 1 , wherein the nucleic acid molecules have a molecular weight of at least about 1×10 10  g/mol, about 1×10 9  g/mol, about 1×10 8  g/mol, about 1×10 7  g/mol, about 1×10 6  g/mol, or about 1×10 5  g/mol. 
     
     
         25 . The particle of  claim 1 , wherein the core has a negative or positive surface charge. 
     
     
         26 . The particle of  claim 1 , further comprising one or more agents for delivery within the core. 
     
     
         27 . The particle of  claim 26 , wherein the agent comprises a chemotherapeutic agent selected from the group consisting of doxorubicin, carboplatin, cisplatin, cyclophosphamide, docetaxel, erlotinib, etoposide, fluorouracil, gemcitabine, imatinib mesylate, irinotecan, methotrexate, paclitaxel, sorafinib, sunitinib, topotecan, vincristine, vinblastine and combination thereof. 
     
     
         28 . The particle of  claim 1 , wherein the first greatest dimension of the core is greater than 2 μm, 1 μm, 500 nm, 200 nm, 100 nm or 50 nm. 
     
     
         29 . The particle of  claim 1 , wherein the second greatest dimension of the core is less than 500 nm, 200 nm, 100 nm, 50 nm, 20 nm or 10 nm. 
     
     
         30 . The particle of  claim 1 , further comprising a film coated on the core, so that the core has its second configuration. 
     
     
         31 . The particle of  claim 30 , wherein the film is or comprises a material selected from the group consisting of an organic material, an inorganic material, or combination thereof. 
     
     
         32 . The particle of  claim 30 , wherein the film is or comprises a polymer. 
     
     
         33 . The particle of  claim 32 , wherein the film is or comprises a lipid. 
     
     
         34 . The particle of  claim 30 , wherein the film is or comprises at least one polyelectrolyte layer. 
     
     
         35 . The particle of  claim 34 , wherein the polyelectrolye layer is degradable or non-degradable. 
     
     
         36 . The particle of  claim 34 , wherein the polyelectrolyte layer is or comprises a polycation or polyanion. 
     
     
         37 . The particle of  claim 36 , wherein the polycation is selected from the group consisting of polyethylenimine, poly(L-lysine) (PLL), poly(lactic acid) (PLA), derivatives and combination thereof. 
     
     
         38 . The particle of  claim 30 , wherein the film comprises a layer-by-layer (LBL) film. 
     
     
         39 . The particle of  claim 38 , wherein the LBL film comprises multiple polyelectrolyte layers. 
     
     
         40 . The particle of  claim 39 , wherein the LBL film comprises multiple polyelectrolyte layers of alternating charges. 
     
     
         41 . The particle of  claim 30 , wherein the film further comprises one or more agents. 
     
     
         42 . The particle of  claim 30 , wherein the coated particle has a positive or negative surface charge. 
     
     
         43 . A particle, comprising:
 a core comprised of one or more nucleic acid molecules self-assembled in a crystalline structure comprising lamellar sheets.   
     
     
         44 . A method for forming a particle comprising:
 assembling one or more nucleic acid molecules into a core with a crystalline structure comprising lamellar sheets.   
     
     
         45 . A method for forming a particle comprising:
 assembling one or more nucleic acid molecules into a core, wherein the core has a first greatest dimension greater than 2 μm, and   coating the core with a film, wherein the coated core has a second greatest dimension less than 500 nm.   
     
     
         46 . The method of  claim 45 , further comprising forming the nucleic acid molecules via rolling circle amplification (RCA), rolling circle transcription (RCT) or both. 
     
     
         47 . The method of  claim 46 , wherein the step of forming comprises using a circular nucleic acid template. 
     
     
         48 . The method of  claim 47 , wherein the step of forming comprises hybridizing the circular nucleic acid template with a primer. 
     
     
         49 . The method of  claim 48 , wherein the primer is complementary to a portion of the circular nucleic acid template. 
     
     
         50 . The method of  claim 47 , wherein the step of forming further comprises amplifying the circular nucleic acid template using an enzyme. 
     
     
         51 . The method of  claim 50 , wherein the enzyme is Φ29 DNA polymerase, T7 polymerase or both. 
     
     
         52 . The method of  claim 45 , wherein the step of coating comprises mixing the core in a coating solution. 
     
     
         53 . The method of  claim 52 , wherein the coating solution comprises polyethylenimine. 
     
     
         54 . The method of  claim 45 , wherein the step of coating further comprises sequentially assembling additional layers. 
     
     
         55 . A method for using a particle:
 administering or implanting a particle of  claim 1  to a subject.   
     
     
         56 . The method of  claim 55 , further comprising exposing the particle to a cleavage agent so that the nucleic acid molecules are cleaved into copies of nucleic acids. 
     
     
         57 . The method of  claim 56 , wherein the cleavage agent is an enzyme selected from the group consisting of nuclease, Dicer, DNAase, RNAase and combination thereof. 
     
     
         58 . The method of  claim 57 , further comprising releasing the cleaved copies of nucleic acids. 
     
     
         59 . The method of  claim 55 , wherein the particles are used for dysregulation of genes.