Methods for Transfecting Nucleic Acid Into Live Cells

Abstract

The present invention includes methods for transferring a multigenic phenotype to a cell by transfecting, preferably by phototransfection, and locally transfecting a cell or a cellular process with a laser while the cell is bathed in a fluid medium comprising two or more nucleic acids, thereby introducing the nucleic acid into the interior of the cell. Expression of the nucleic acids results in a multigenic phenotype in the transfected cell.

1 - 25 . (canceled) 
     
     
         26 . A method of transferring a multigenic phenotype to a recipient cell, the method comprising: transfecting a recipient cell with at least two nucleic acids, wherein the nucleic acids enter the recipient cell, wherein the nucleic acids are functional in the recipient cell and wherein the multigenic phenotype is transferred to the recipient cell. 
     
     
         27 . The method of  claim 26 , wherein the recipient cell is a eukaryotic cell. 
     
     
         28 . The method of  claim 26 , wherein the recipient cell is a mammalian cell. 
     
     
         29 . The method of  claim 26 , wherein the recipient cell is selected from the group consisting of: an epithelial cell, a neuron, a fibroblast, an embryonic fibroblast, a keratinocyte, an adult stem cell, an embryonic stem cell, an astrocyte and a cardiomyocyte. 
     
     
         30 . The method of  claim 26 , wherein the at least two nucleic acids are selected from the group consisting of: an RNA, a DNA and a combination thereof. 
     
     
         31 . The method of  claim 30 , wherein the RNA is at least one selected from the group consisting of: mRNA, siRNA, miRNA, hnRNA, and tRNA. 
     
     
         32 . The method of  claim 30 , wherein the RNA is a mRNA. 
     
     
         33 . The method of  claim 32 , wherein the mRNA is expressed by the recipient cell. 
     
     
         34 . The method of  claim 32 , wherein the mRNA is isolated from a donor cell. 
     
     
         35 . The method of  claim 34 , wherein the donor cell is selected from the group consisting of: an astrocyte, a cardiomyocyte, a neonatal cardiomyocyte, an embryonic stem cell and a neuron. 
     
     
         36 . The method of  claim 32 , wherein the mRNA is in vitro transcribed. 
     
     
         37 . The method of  claim 32 , wherein the nucleic acids are prepared by a method selected from the group consisting of: mRNA isolation from a donor cell, in vitro mRNA transcription or mRNA chemical synthesis. 
     
     
         38 . The method of  claim 26 , wherein the nucleic acids comprise at least two different mRNAs encoding at least three different polypeptides. 
     
     
         39 . The method of  claim 26 , wherein the nucleic acids comprise at least three different mRNAs encoding at least three different polypeptides. 
     
     
         40 . The method of  claim 26 , wherein the nucleic acids comprise at least four different mRNAs encoding at least four different polypeptides. 
     
     
         41 . The method of  claim 26 , wherein the nucleic acids comprise at least five different mRNAs encoding at least five different polypeptides.