METHOD FOR DETERMINING DESIRED CELL TYPE USING EXPRESSION OF miRNA AS INDICATOR

Abstract

A method is provided for distinguishing living cells in a living state with high accuracy. Provided is a method for distinguishing a desired cell type from a cell group comprising two or more types of cells, using the expression of miRNA as an indicator, wherein the method comprises the following steps: (1) a step of introducing mRNA comprising a marker gene operably linked to the target sequence of miRNA used as an indicator into a cell group; and (2) a step of distinguishing a cell type, using the translation level of the marker gene as an indicator.

What is claimed is: 
     
         1 . A method for distinguishing a desired cell type from a cell group comprising two or more types of cells, using the expression of miRNA as an indicator, wherein the method comprises the following steps:
 (1) a step of introducing mRNA comprising a marker gene operably linked to a target sequence of miRNA used as an indicator into a cell group; and   (2) a step of distinguishing a cell type, using the translation level of the marker gene as an indicator.   
     
     
         2 . The method according to  claim 1 , wherein the step of introducing mRNA is carried out by an electroporation method. 
     
     
         3 . The method according to  claim 1 , wherein the target sequence of the miRNA is linked to the 5′- or 3′-terminal side of the marker gene. 
     
     
         4 . The method according to  claim 3 , wherein the step of introducing mRNA is carried out by an electroporation method. 
     
     
         5 . A method for producing an mRNA comprising a marker gene operably linked to a target sequence of miRNA used as an indicator, comprising:
 preparing a template DNA containing a promoter sequence; and   synthesizing the mRNA from the template DNA using an in vitro transcription synthesis method.   
     
     
         6 . The method according to  claim 5 , further comprising step of designing the mRNA containing 5′-UTR, a coding region of the marker gene and 3′-UTR, in which target sequence of miRNA is contained in the 5′-UTR or 3′-UTR, and
 wherein the step of preparing the template DNA further comprises: 
 preparing a template of the 5′-UTR, the coding region of the marker gene and the 3′-UTR; and 
 linking the template of the 5′-UTR, the coding region of the marker gene and the 3′-UTR using a PCR primer.