BIOLOGICALLY ACTIVE SYNTHETIC NANOPARTICLE CONSTRUCTS AND METHODS OF USE THEREOF
This application discloses the compositions comprising biologically active synthetic nanoparticle constructs and methods of use thereof to modify gene expression including transcriptional activation and transcriptional repression.
1 . A method of transcribing a target gene in a cell to produce a peptide encoded by the target gene, comprising delivering to the cell a biologically active synthetic nanoparticle construct comprising a biologically inert substrate on which there is covalently bonded a plurality of DNA binding domains, a plurality of nuclear localization signals and a plurality of activation domains, wherein the plurality of DNA binding domains binds to a target DNA sequence and wherein the plurality of activation domains recruits endogenous transcriptional basal machinery.
2 . The method of claim 1 wherein the transcribing of the target gene in the cell occurs in a nucleus.
3 . The method of claim 1 wherein the cell comprises a stem cell.
4 . The method of claim 1 wherein the target gene comprises a gene regulating cellular differentiation.
5 . The method of claim 1 wherein the target DNA sequence is an enhancer region.
6 . The method of claim 1 wherein the biologically inert substrate is selected from the group consisting of: magnetic nanoparticles, magnetic-core shell nanoparticles, silica nanoparticles, mesoporous nanoparticles, quantum dots, supramolecular nanoparticles, and polymer-based nanoparticles.
7 . The method of claim 1 wherein the biologically inert substrate is a gold nanoparticle.
8 . The method of claim 1 where the plurality of DNA binding domains comprises hairpin polyamides.
9 . The method of claim 9 wherein the hairpin polyamides comprise at least one N-methyl-imidazole moiety or at least one N-methyl pyrrole moiety, or combinations thereof, arranged sequentially on said polyamide to bind a target gene.
10 . The method of claim 1 wherein the plurality of DNA binding domains is selected from the group consisting of zinc finger domains, triple forming oligonucleotides, transcription activator-like effectors, oligonucleotide analogs, locked-nucleic acids, and peptide nucleic acids, and combinations thereof.
11 . The method of claim 1 wherein the plurality of DNA binding domains is attached to a surface of the biologically inert substrate by crosslinking molecules.
12 . The method of claim 1 wherein the plurality of nuclear localization signals is attached to the surface of the biologically inert substrate by crosslinking molecules.
13 . The method of claim 1 wherein the plurality of activation domains is attached to a surface of the biologically inert substrate by crosslinking molecules.
14 . The method of claim 1 wherein the plurality of nuclear localization signals is monopartite.
15 . The method of claim 1 , wherein the plurality of nuclear localization signals is bipartite.
16 . The method of claim 1 wherein the plurality of nuclear localization signals is derived from a SV-40 antigen.
17 . The method of claim 1 wherein the plurality of nuclear localization signals is derived from a HIV-1 antigen.
18 . The method of claim 1 wherein the plurality of nuclear localization signals is derived from the group consisting of TAT, Penetratin, MAP, Transportin/TP10, VP22, MPG, Pep1, pVEC, YTA2, YTA4, M918, and CADY, and combinations thereof.
19 . The method of claim 1 wherein the plurality of activation domains is selected from the group consisting of peptoids, amphipathic isoxasolidine, wrenchnolol, and amphipathic helix peptides, and combinations thereof.
20 . A method of repressing transcription of a target gene in a cell comprising delivering to the cell a biologically active synthetic nanoparticle construct comprising a biologically inert substrate on which there is covalently bonded a plurality of DNA binding domains, a plurality of nuclear localization signals and a plurality of repression domains, wherein the plurality of DNA binding domains binds to a target DNA sequence.