Human induced pluripotent stem cells for live cell cycle monitoring and endogenous gene activation
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Summary
This summary is machine-generated.We developed novel human induced pluripotent stem cell (hiPSC) lines for simultaneous monitoring of cell cycle progression and gene activation. These Myo-CCER and CraCCER lines enable advanced research in stem cell biology and muscle development.
Area Of Science
- Stem cell biology
- Cell cycle regulation
- Gene editing technologies
Background
- Fluorescence Ubiquitination Cell Cycle Indicator (FUCCI) is used to track cell cycle in live cells, including human induced pluripotent stem cells (hiPSC).
- Previous work established hiPSC with dCas9VPR for gene activation and ACTN2-Citrine for muscle cell monitoring.
Purpose Of The Study
- To create dual and triple transgenic hiPSC lines integrating FUCCI with or without dCas9VPR into existing ACTN2-Citrine lines.
- To validate the functionality of these novel transgenic hiPSC lines for simultaneous cell cycle and gene expression studies.
Main Methods
- Genomic integration of FUCCI and dCas9VPR transgenes into ROSA26 and AAVS1 loci of hiPSC lines.
- Utilizing established ACTN2-Citrine reporter lines for muscle cell development tracking.
- Demonstration of transgene functionality in the newly developed hiPSC lines.
Main Results
- Successful generation of dual and triple transgenic hiPSC lines, named Myo-CCER and CraCCER.
- Confirmed functionality of integrated FUCCI and dCas9VPR systems within the hiPSC lines.
- Demonstrated utility for monitoring both cell cycle and gene expression/sarcomere development.
Conclusions
- The novel Myo-CCER and CraCCER hiPSC lines provide a powerful platform for studying cell cycle dynamics alongside gene activation and muscle development.
- These advanced hiPSC models facilitate multi-parametric analysis in stem cell research.
- Enables simultaneous tracking of cell cycle and specific cellular processes in hiPSC-derived cells.
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