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A Novel Feeder-free System for Mass Production of Murine Natural Killer Cells In Vitro
Published on: January 9, 2018
Target cell-induced perforin mRNA turnover in NK3.3 cells is mediated by multiple elements within the mRNA coding
W S Goebel1, R H Schloemer, Z Brahmi
1Department of Microbiology, Indiana University School of Medicine, Indianapolis 46202, USA.
Molecular Immunology
|March 1, 1996
Summary
Cytolytic cells rapidly degrade perforin mRNA when encountering sensitive targets. This accelerated mRNA decay, specific to cytotoxic proteins, is regulated by elements within the perforin coding region, not its 3' UTR.
Area of Science:
- Molecular Biology
- Immunology
- Cell Biology
Background:
- Cytolytic cells eliminate target cells through cytotoxic proteins like perforin.
- Perforin mRNA levels are known to decrease upon exposure to sensitive targets.
Purpose of the Study:
- To investigate the mechanism of perforin mRNA downregulation in NK3.3 cells upon exposure to target cells (TC).
- To identify the specific regions within the perforin transcript responsible for target cell-mediated mRNA turnover.
Main Methods:
- Exposure of NK3.3 cells to sensitive TC to induce perforin mRNA decay.
- Measurement of perforin and beta-actin mRNA levels.
- Cloning of perforin cDNA segments into the 3' UTR of rabbit beta globin (RG) constructs.
- Analysis of mRNA turnover rates in engineered constructs.
Main Results:
- Perforin mRNA undergoes accelerated turnover in NK3.3 cells exposed to TC.
- This decay is specific to cytotoxic protein messages, as beta-actin mRNA levels remain unchanged.
- The TC-induced turnover is not due to inhibited RNA synthesis or a short intrinsic mRNA half-life.
- Elements within the perforin coding region, not the 3' UTR, mediate TC-induced mRNA turnover.
Conclusions:
- Target cell exposure induces a novel mRNA decay mechanism for perforin.
- Regulatory elements controlling perforin mRNA stability are located within its coding region.
- This finding represents a previously undescribed mode of mRNA regulation.
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