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Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
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In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing...
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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
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The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
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An Efficient Method for Directed Hepatocyte-Like Cell Induction from Human Embryonic Stem Cells
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Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells.

Sangtae Yoon1,2, Kyojin Kang1,2, Young-Duck Cho3

  • 1HY Indang Center of Regenerative Medicine and Stem Cell Research, Hanyang University, Seoul 04763, Republic of Korea.

Biomed Research International
|October 18, 2018
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Summary
This summary is machine-generated.

Researchers developed a novel, non-integrating method using mRNA to directly convert fibroblasts into hepatocyte-like cells (R-iHeps), offering potential for cell therapy without genomic risks.

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Area of Science:

  • Cell Biology
  • Regenerative Medicine
  • Hepatology

Background:

  • Direct reprogramming bypasses pluripotent stages for cell differentiation.
  • Viral vectors used in reprogramming pose genome integration risks.

Purpose of the Study:

  • To develop a non-integrating direct reprogramming method for generating hepatocyte-like cells.
  • To assess the potential clinical applications of mRNA-based cell conversion.

Main Methods:

  • Transfection of mouse embryonic fibroblasts with mRNA encoding Foxa3 and HNF4α.
  • Culturing converted cells to form hepatocyte-like colonies (R-iHeps).
  • Assessing R-iHeps for hepatocyte-specific markers, function, and in vivo engraftment.

Main Results:

  • Fibroblasts converted to epithelial-morphology hepatocyte-like cells (R-iHeps) within 10-12 days.
  • R-iHeps expressed key hepatocyte markers (albumin, HNF4α, CK18, CYP1A2) and exhibited hepatic functions.
  • Successful engraftment and FAH enzyme expression were confirmed in mouse models.

Conclusions:

  • Non-integrating mRNA-based direct reprogramming is a viable method for generating functional hepatocyte-like cells.
  • This approach avoids genomic integration risks associated with viral vectors.
  • The developed method shows promise for future cell therapy applications in liver diseases.