Method for determining desired cell type using expression of miRNA as indicator
AssigneeKyoto University
InventorsHirohide Saito, Kei Endo
Life of this patent
Estimating the term…
Abstract
A method for distinguishing living cells in a living state with high accuracy. 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.
Claims24
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 a reporter mRNA comprising a Cap structure or Cap analogue, a nucleic acid sequence encoding a marker protein operably linked to a target sequence of miRNA into a cell group, and a polyA tail, wherein the reporter mRNA is introduced into the cell group in a form of a synthetic RNA molecule without using a virus vector construct or DNA vector construct, and wherein the target sequence of miRNA is linked to the 5′-terminal side of the nucleic acid sequence encoding the marker protein;
- and
- (2) a step of distinguishing the desired cell type, using a translation level of the marker protein.
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 simultaneously introducing two or more reporter mRNAs into a cell group, wherein the two or more reporter mRNAs comprise a Cap structure or Cap analogue, a nucleic acid sequence encoding a marker protein gene operably linked to a target sequence of miRNA and a poly A tail;
- wherein the target sequence of miRNA is linked to the 5′-terminal side of the nucleic acid sequence encoding the marker protein;
- and wherein the two or more reporter mRNAs each comprise a different target sequence and a different nucleic acid sequence encoding a different marker protein, and the two or more mRNAs are introduced into the cell group in a form of a synthetic RNA molecule without using a virus vector construct or DNA vector construct;
- and
- (2) a step of distinguishing the desired cell type, using a translation level of the marker protein.
A kit for distinguishing a cell, which comprises:
- (a) one or more reporter mRNA comprising a Cap structure or Cap analogue, a nucleic acid sequence encoding a marker protein operably linked to a target sequence of miRNA and a polyA tail, wherein the target sequence of miRNA is linked to the 5′-terminal side of the nucleic acid sequence encoding the marker protein, with the proviso that when two or more reporter mRNAs are present, the two or more reporter mRNAs each comprise a different target sequence and a different nucleic acid sequence encoding a different marker protein;
- and
- (b) a control mRNA comprising a Cap structure or Cap analogue, a nucleic acid sequence encoding a control marker protein and a polyA tail, wherein the control mRNA does not include a target sequence of miRNA and the control marker protein is different from the marker protein of the reporter mRNA.
A kit for distinguishing a cell, which comprises:
- two or more reporter mRNAs comprising a Cap structure or Cap analogue, a nucleic acid sequence encoding a marker protein operably linked to a target sequence of miRNA and a polyA tail, wherein the target sequence of miRNA is linked to the 5′-terminal side of the nucleic acid sequence encoding the marker protein, and wherein the two or more reporter mRNAs each comprise a different target sequence and a different nucleic acid sequence encoding a different marker protein.
A kit for distinguishing a cell, which comprises:
- one or more reporter mRNA comprising a Cap structure or Cap analogue, a nucleic acid sequence encoding a marker protein operably linked to a target sequence of miRNA, and a polyA tail, wherein the target sequence of miRNA is linked to the 5′-terminal side of the nucleic acid sequence encoding the marker protein.
Dependencies are inferred from the claim text.