Using IL-12 p40 as a Reporter Gene in Stem Cell Research
Author Information
Author(s): D'Aiuto Leonardo, Robison Clinton S, Gigante Margherita, Nwanegbo Edward, Shaffer Benjamin, Sukhwani Meena, Castro Carlos A, Chaillet J Richard
Primary Institution: University of Pittsburgh School of Medicine
Hypothesis
Can human IL-12 p40 be effectively used as a reporter gene for high-throughput screening of engineered mouse embryonic stem cells?
Conclusion
IL-12 p40 is a sensitive and effective reporter gene that does not compromise the pluripotency of mouse embryonic stem cells.
Supporting Evidence
- IL-12 p40 was first detected in the medium of transfected cells 12 hours after transfection.
- The maximum IL-12 p40 concentration was observed 72 hours after transfection.
- The amount of IL-12 p40 secreted increased with the amount of plasmid transfected up to 1 μg DNA.
- IL-12 p40 expression was proportional to the amount of mRNA present in the cells.
- The ELISA procedure identified 50% of rtTA-containing clones expressing IL-12 p40.
Takeaway
Researchers found that a protein called IL-12 p40 can help scientists quickly find modified stem cells without hurting their ability to grow and change into different cell types.
Methodology
The study involved transfecting BHK-21 and COS-7 cells with plasmids containing IL-12 p40 and measuring its expression using ELISA.
Limitations
The study does not address the versatility of IL-12 p40 as a reporter gene across all applications.
Digital Object Identifier (DOI)
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