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Related Concept Videos

Methods of Nuclear Reprogramming01:24

Methods of Nuclear Reprogramming

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Nuclear reprogramming is a process of transforming one cell type into an unrelated cell type by epigenetic changes that alter the cell’s original gene expression pattern. Such epigenetic changes force cells to express a different set of genes, which play a significant role in inducing transformation into other cell types. Nuclear reprogramming offers applications in reproductive cloning for livestock propagation and regenerative medicine — developing patient-specific cells for...
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Reprogramming alters the gene expression in somatic cells, transforming them into induced pluripotent stem (iPS) cells over several generations. Scientists can reprogram cells by introducing genes for four transcription factors—Oct4, Sox2, Klf4, and c-Myc (OSKM) by viral or non-viral methods. These factors are also known as Yamanaka factors after Shinya Yamanaka, who first generated iPS cells using mouse skin cells. Yamanaka was awarded the Nobel Prize in Physiology or Medicine in 2012...
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Introduction to Nuclear Reprogramming01:14

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Nuclear reprogramming is the process of switching gene expression of one cell type to that of another cell type, usually from a differentiated cell state to an undifferentiated cell state. Differentiation occurs during processes such as development and morphogenesis, tissue regeneration, and malignancy. Cells can also be artificially induced to reprogram their gene expression by techniques such as nuclear transfer, induced pluripotency, and cell fusion. Such techniques have many applications in...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Deterministic transfection drives efficient nonviral reprogramming and uncovers reprogramming barriers.

Daniel Gallego-Perez1, Jose J Otero2, Catherine Czeisler3

  • 1Department of Chemical and Biomolecular Engineering, College of Engineering, The Ohio State University, Columbus, OH; Department of Surgery, College of Medicine, The Ohio State University, Columbus, OH; Center for Regenerative Medicine and Cell-Based Therapies (CRMCBT), The Ohio State University, Columbus, OH.

Nanomedicine : Nanotechnology, Biology, and Medicine
|December 30, 2015
PubMed
Summary
This summary is machine-generated.

A novel nanotechnology platform enables efficient and controlled cell reprogramming for regenerative medicine. This non-viral method achieves high reprogramming efficiencies, overcoming limitations of current viral and non-viral techniques for clinical applications.

Keywords:
Induced neuronNanochannel electroporationNuclear reprogrammingTransfection

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Area of Science:

  • Cell Biology
  • Regenerative Medicine
  • Nanotechnology

Background:

  • Current nuclear reprogramming methods face safety and consistency challenges, hindering clinical use.
  • Direct neuronal reprogramming is a key area in regenerative medicine for cell fate control.

Purpose of the Study:

  • To introduce a novel non-viral nanotechnology-based platform for deterministic, large-scale cell transfection.
  • To demonstrate the platform's efficacy in direct neuronal reprogramming using transcription factors Brn2, Ascl1, and Myt1l (BAM).

Main Methods:

  • Development of a nanotechnology platform for non-viral, high-resolution cell transfection.
  • Application of the platform for direct neuronal reprogramming via overexpression of BAM transcription factors.
  • High-throughput nanoelectroporation (NEP) for process interrogation.

Main Results:

  • Achieved reprogramming efficiencies comparable to viral methods (up to 9-12%) without capsid size limitations.
  • Demonstrated superior performance over existing non-viral reprogramming techniques.
  • Identified Ascl1 dosage and CCNA2 as key regulators of BAM-mediated reprogramming.
  • Showed that neuronal complexity can be tailored by adjusting the BAM ratio and including additional genes.

Conclusions:

  • The novel nanotechnology platform offers a safe and controllable alternative for nuclear reprogramming.
  • This technology facilitates precise control over cell fate and neuronal complexity.
  • The findings provide a foundation for advancing regenerative medicine and clinical translation of reprogramming.