Solid surface for biomolecule delivery and high-throughput assay

Abstract

The present invention is related to a method for introducing biomolecules, such as nucleic acids, into cells by culturing cells on a solid surface which is coated with a transfection reagents and biomolecules.

What is claimed is:  
     
         1 . A method for introducing oligonucleotides or polypeptides into eukaryotic cells comprising: (a) providing a solid surface at least partially coated with a cationic polymer or a lipid, (b) adding the oligonucleotides or polypeptides to be introduced into the eukaryotic cells onto the solid surface, (c) seeding cells on the solid surface at a sufficient density and under appropriate conditions for introduction of the oligonucleotides or polypeptides into the eukaryotic cells.  
     
     
         2 . The method of  claim 1 , wherein the surface is selected from the group consisting of flasks, dishes, multi-well plates, glass slides, and implanted devices.  
     
     
         3 . The method of  claim 1 , wherein said solid surface is coated with a lipid-polymer.  
     
     
         4 . The method of  claim 1 , wherein said polymer or lipid contain an additional reagent selected from the group consisting of a cell-targeting moiety, an intracellular targeting moiety, a membrane-destabilizing component, and one or more transfection enhancers.  
     
     
         5 . The method of  claim 1 , wherein said cationic polymer or lipid is affixed on the surface by evenly spreading the reagent on the surface or spotting said biomolecule delivery reagent in discrete areas of the surface.  
     
     
         6 . The method of  claim 1 , wherein the cationic polymer or lipid contains a matrix reagent selected from the group consisting of proteins, glycoproteins, peptides, polysaccharides, and polymers or mixtures thereof.  
     
     
         7 . The method of  claim 6 , wherein said protein is selected from gelatin, collagen, laminin, fibronectin, and bovine serum albumin or a mixture thereof.  
     
     
         8 . The method of  claim 6 , wherein said polymer is selected from hydrogels, biodegradable polymers, and biocompatible materials.  
     
     
         9 . The method of  claim 1 , wherein said cationic polymer or lipid contains a cell culture reagent selected from the group consisting of cytoreductive reagents, cell binding reagents, cell growing reagents, cell stimulating reagents, and cell inhibiting reagents.  
     
     
         10 . The method of  claim 1 , wherein the solid surface is selected from a slide or a multi-well plate.  
     
     
         11 . The method of  claim 1 , wherein the eukaryotic cells are mammalian cells.  
     
     
         12 . The method of  claim 11 , wherein the mammalian cells are dividing cells or non-dividing cells.  
     
     
         13 . The method of  claim 11 , wherein the mammalian cells are transformed cells or primary cells.  
     
     
         14 . The method of  claim 11 , wherein the mammalian cells are somatic cells or stem cells.  
     
     
         15 . The method of  claim 1 , wherein the eukaryotic cells are plant cells.  
     
     
         16 . The method of  claim 1 , wherein the cationic polymer or lipid is affixed on the surface by evenly spreading the reagent on the solid surface or spotting the transfection regent on the solid surface manually or by an automated mechanism.  
     
     
         17 . The method of  claim 1 , wherein the oligonucleotide is selected from the group consisting of DNA, RNA, and DNA/RNA hybrid.  
     
     
         18 . The method of  claim 1 , wherein the oligonucleotide is selected from the group consisting of a linear molecule, a plasmid, and single stranded oligodeoxynucleotide (ODN).  
     
     
         19 . The method of  claim 1 , wherein the oligonucleotide is selected from the group consisting of single stranded RNA (ribozyme) or double stranded RNA (siRNA).  
     
     
         20 . The method of  claim 1 , wherein the eukaryotic cells are insect cells.  
     
     
         21 . A method of determining whether a biomolecule can enter a cell comprising (a) providing a solid surface coated with a polymer or lipid to which said biomolecule can interact, (b) adding the biomolecules to the solid surface such that the biomolecules interact with said polymer or lipid, (c) seeding cells onto the surface with sufficient density and under appropriate conditions for introduction of the biomolecules into the cells, and (d) detecting whether the biomolecule has been delivered to the cells.  
     
     
         22 . The method of  claim 21  wherein the biomolecules are selected from the group consisting of nucleotides, proteins, peptides, sugars, polysaccharides, and organic compounds.  
     
     
         23 . The method of  claim 22  wherein the oligonucleotides are selected from the group consisting of DNA, RNA, and DNA/RNA hybrid.  
     
     
         24 . The method of  claim 23  wherein the oligonucleotides are selected from the group consisting of a linear molecule, a plasmid, and single stranded oligodeoxynucleotide (ODN).  
     
     
         25 . The method of  claim 23 , wherein the oligonucleotides are selected from the consisting of single stranded RNA (ribozyme) or double stranded RNA (siRNA).  
     
     
         26 . The method of  claim 21 , wherein the eukaryotic cells are mammalian cells.  
     
     
         27 . The method of  claim 26 , wherein the mammalian cells are dividing cells or non-dividing cells.  
     
     
         28 . The method of  claim 26 , wherein the mammalian cells are transformed cells or primary cells.  
     
     
         29 . The method of  claim 26 , wherein the mammalian cells are somatic cells or stem cells.  
     
     
         30 . The method of  claim 21 , wherein the cells are selected from the group consisting of plant, insect, and bacterial cells.  
     
     
         31 . The method of  claim 1 , wherein the cationic polymer or lipid is affixed on the surface manually or by an automated mechanism.  
     
     
         32 . The method of  claim 21 , wherein said detecting is performed by detecting said biomolecule, a product of the biomolecule, activity of a biomoloecule, activity of the product of the biomolecule, a target molecule, a product catalyzed by the biomolecule or a product regulated by the biomolecule.