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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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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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Reproductive cloning is the process of producing a genetically identical copy—a clone—of an entire organism. While clones can be produced by splitting an early embryo—similar to what happens naturally with identical twins—cloning of adult animals is usually done by a process called somatic cell nuclear transfer (SCNT).
Somatic Cell Nuclear Transfer
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Genetic transfer occurs when genetic information is passed from one organism to another. It occurs via two mechanisms: vertical gene transfer and horizontal gene transfer. Vertical gene transfer occurs when genetic information is transferred from one generation to the next, which happens much more frequently than horizontal gene transfer. Both sexual and asexual reproduction are forms of vertical gene transfer, where one or more organisms pass some or all of their genome onto their progeny.
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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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The first successfully cloned mammal was Dolly, a sheep, born on 5th July 1996 at Roslin Institute, Scotland. The cloned sheep was named after the American singer Dolly Parton. Dolly lived for seven years and died of respiratory complications, which is speculated to be due to the actual age of her DNA. Because the DNA in cloned cells belongs to an older individual,  the cloned individual’s life expectancy may be affected. Indeed, analysis of Dolly’s DNA revealed shorter...
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Bovid Interspecies Somatic Cell Nuclear Transfer with Ooplasm Transfer.

L Antonio González-Grajales1, Gabriela F Mastromonaco2

  • 1Institut für Fortpflanzung landwirtschaftlicher Nutztiere Schönow e.V, Bernau bei Berlin, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|April 11, 2023
PubMed
Summary

Interspecies somatic cell nuclear transfer with ooplasm transfer (iSCNT-OT) can help conserve endangered species. This method combines bison cells and oocyte cytoplasm into cow eggs, potentially overcoming nuclear-mitochondrial incompatibilities.

Keywords:
CloningCross-speciesCybridCytoplasm transferHeteroplasmyInterspeciesMicromanipulationNuclear transplantation

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Area of Science:

  • Reproductive biology
  • Conservation genetics
  • Cellular and molecular biology

Background:

  • Interspecies somatic cell nuclear transfer (iSCNT) is vital for endangered species preservation.
  • Nuclear-mitochondrial incompatibilities limit iSCNT's effectiveness.
  • Ooplasm transfer (iSCNT-OT) offers a potential solution to these incompatibilities.

Purpose of the Study:

  • To develop and describe an iSCNT-OT protocol for interspecies applications.
  • To investigate the potential of iSCNT-OT in overcoming nuclear-mitochondrial communication barriers.
  • To establish a foundation for future research on interspecies embryonic development.

Main Methods:

  • A novel two-step electrofusion protocol was employed.
  • Bison (Bison bison bison) somatic cells and oocyte cytoplasm were transferred.
  • The components were introduced into enucleated bovine (Bos taurus) oocytes.

Main Results:

  • The study successfully describes a combined iSCNT-OT procedure.
  • The protocol integrates bison nuclear and ooplasmic components into bovine oocytes.
  • This establishes a model for studying interspecies nuclear-mitochondrial interactions.

Conclusions:

  • The described iSCNT-OT protocol demonstrates a viable approach for interspecies applications.
  • This technique holds promise for overcoming reproductive barriers in conservation efforts.
  • Further research can utilize this method to explore nuclear-cytoplasmic crosstalk in hybrid embryos.