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Related Concept Videos

Embryonic Stem Cells00:58

Embryonic Stem Cells

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Embryonic stem (ES) cells are undifferentiated pluripotent cells, meaning they can produce any cell type in the body. This gives them tremendous potential in science and medicine since they can generate specific cell types for use in research or to replace body cells lost due to damage or disease.
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Stem cell research aims to find ways to use stem cells to regenerate and repair cellular damage. Over time, most adult cells undergo the wear and tear of aging and lose their ability to divide and repair themselves. Stem cells do not display a particular morphology or function. Adult stem cells, which exist as a small subset of cells in most tissues, keep dividing and can differentiate into a number of specialized cells generally formed by that tissue. These cells enable the body to renew and...
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Embryonic stem (ES) cells were first discovered in mice in 1981 by Martin Evans. In 1998, James Thomson identified a method to isolate embryonic stem cells from humans. Human embryonic stem cells (hESCs) are obtained from 3-5 day old embryos that remain unused after an in vitro fertilization procedure.
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Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells
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Targeting embryonic stem cells.

Roland H Friedel1

  • 1Helmholtz Center Munich, Institute of Developmental Genetics, Neuherberg, Germany. roland.friedel@helmholtz-muenchen.de

Methods in Molecular Biology (Clifton, N.J.)
|June 9, 2009
PubMed
Summary
This summary is machine-generated.

Gene targeting in mouse embryonic stem cells allows precise genome engineering. This method uses electroporation and drug selection to identify correctly modified cells for creating genetically engineered mice.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Developmental Biology

Background:

  • Mouse embryonic stem cells are crucial for genetic engineering.
  • Precise modification of the mouse genome is essential for studying gene function.

Purpose of the Study:

  • To outline a robust method for gene targeting in mouse embryonic stem cells.
  • To detail the process from vector introduction to generating chimeric mice.

Main Methods:

  • Gene targeting vector introduction via electroporation.
  • Drug selection to isolate modified cell clones.
  • Screening for homologous recombination using Southern blot or long-range PCR.

Main Results:

  • Successful isolation of correctly targeted embryonic stem cell clones.
  • Validation of recombination events through molecular analysis.
  • Generation of chimeric mice from selected clones.

Conclusions:

  • Gene targeting in mouse embryonic stem cells is an effective strategy for genome engineering.
  • The described methods ensure reliable identification and expansion of desired clones.
  • This approach facilitates the creation of genetically modified mouse models.