miRNA-Responsive mRNA for Skeletal Muscle Progenitor Cell Sorting
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Solution Overview
Problem
Current methods for purifying skeletal muscle progenitor cells from pluripotent stem cells are inefficient due to the lack of specific cellular surface antigens, particularly for human skeletal muscle progenitor cells, which hinders their utilization in cell transplantation therapy.
Innovation Solution
A method involving the introduction of miRNA-responsive mRNA into cell populations, specifically using sequences recognized by miRNAs like miR-1, miR-133, and miR-206 to express marker proteins, allowing for the efficient sorting of skeletal muscle progenitor cells based on differential expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If antibody-based purification using cellular surface antigens is used, then cell purification can be achieved, but human skeletal muscle progenitor cells cannot be purified because they lack appropriate cellular surface antigens
Solution Approach 1:
The patent introduces an intermediary system consisting of miRNA-responsive mRNA and miRNA-specific antibodies. The miRNA-responsive mRNA acts as a mediator that translates the internal miRNA expression profile into an external detectable marker (marker protein expression), which can then be targeted by antibodies for purification. This intermediary system bridges the gap between the lack of natural surface antigens and the need for antibody-based purification.
Solution Approach 2:
The patent changes the parameter being measured from direct cellular surface antigen presence to marker protein expression levels controlled by miRNA-responsive elements. By shifting from detecting native surface antigens to detecting induced marker proteins whose translation is suppressed by specific miRNAs, the method becomes applicable to human skeletal muscle progenitor cells that previously lacked suitable surface markers.
2Productivity
If miRNA-responsive mRNA is introduced to enable sorting, then cell sorting efficiency improves, but the system complexity increases due to additional molecular components
Solution Approach 1:
The miRNA-responsive mRNA system is self-regulating through endogenous miRNAs. The introduced mRNA contains sequences that are naturally recognized and regulated by the cell's own miRNA profile, eliminating the need for external control mechanisms. The system automatically differentiates skeletal muscle progenitor cells from other cells based on their intrinsic miRNA expression patterns, achieving self-service purification.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method enables the selective isolation of skeletal muscle progenitor cells with high purity, suitable for transplantation, by controlling the translation of marker proteins in response to specific miRNAs, facilitating their identification and separation.
Implementation Method 1
the nucleic acid of (ii) is functionally linked to the nucleic acid of (i) such that the translation amount of the marker protein is suppressed by inhibited translation of the mRNA in response to the miRNA
Data Source
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AI summary
The present invention aims to provide a method of sorting a skeletal muscle progenitor cell from a cell population containing the skeletal muscle progenitor cell. The above-mentioned problem is solved by providing a step of introducing miRNA-responsive mRNA into a cell population. The miRNA-responsive mRNA contains (i) a nucleic acid having a sequence specifically recognized by miRNA specifically expressed in a skeletal muscle progenitor cell, and (ii) a nucleic acid containing a sequence encoding a marker protein.