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Related Concept Videos

Gene Evolution - Fast or Slow?02:05

Gene Evolution - Fast or Slow?

The genomes of eukaryotes are punctuated by long stretches of sequence which do not code for proteins or RNAs. Although some of these regions do contain crucial regulatory sequences, the vast majority of this DNA serves no known function. Typically, these regions of the genome are the ones in which the fastest change, in evolutionary terms, is observed, because there is typically little to no selection pressure acting on these regions to preserve their sequences.
In contrast, regions which code...
Non-equilibrium in the Cell01:16

Non-equilibrium in the Cell

An important concept in studying metabolism and energy is that of chemical equilibrium. Most chemical reactions are reversible. They can proceed in both directions, releasing energy into their environment in one direction, and absorbing it from the environment in the other direction. The same is true for the chemical reactions involved in cell metabolism, such as the breaking down and building up of proteins into and from individual amino acids, respectively. Reactants within a closed system...
Gene Evolution - Fast or Slow?02:05

Gene Evolution - Fast or Slow?

The genomes of eukaryotes are punctuated by long stretches of sequence which do not code for proteins or RNAs. Although some of these regions do contain crucial regulatory sequences, the vast majority of this DNA serves no known function. Typically, these regions of the genome are the ones in which the fastest change, in evolutionary terms, is observed, because there is typically little to no selection pressure acting on these regions to preserve their sequences.
In contrast, regions which code...
Introduction to Nuclear Reprogramming01:14

Introduction to Nuclear Reprogramming

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...
Somatic to iPS Cell Reprogramming01:29

Somatic to iPS Cell Reprogramming

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 for this...
Methods of Nuclear Reprogramming01:24

Methods of Nuclear Reprogramming

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 injury repair.

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A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
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Programming cells by multiplex genome engineering and accelerated evolution.

Harris H Wang1,2,3, Farren J Isaacs1, Peter A Carr4,5

  • 1Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.

Nature
|July 28, 2009
PubMed
Summary
This summary is machine-generated.

Multiplex automated genome engineering (MAGE) rapidly creates vast genomic diversity in cells. This technology accelerates the evolution of organisms for improved industrial applications, like enhanced lycopene production.

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Area of Science:

  • Synthetic Biology
  • Genomics
  • Metabolic Engineering

Background:

  • Generating genomic diversity in the lab is challenging for practical timescales.
  • Existing directed evolution methods are laborious and limited to single-gene manipulation.
  • There is a need for parallel and continuous evolution of gene networks and genomes.

Purpose of the Study:

  • To describe multiplex automated genome engineering (MAGE) for large-scale cell programming and evolution.
  • To develop automated devices for rapid and continuous generation of diverse genetic changes.
  • To optimize the DXP biosynthesis pathway for overproduction of lycopene.

Main Methods:

  • MAGE simultaneously targets multiple genomic locations within single cells or populations.
  • Automated MAGE devices facilitate cyclical and scalable generation of combinatorial genomic diversity.
  • Simultaneous modification of 24 genetic components in the DXP pathway using synthetic DNA pools.

Main Results:

  • Generated over 4.3 billion combinatorial genomic variants per day.
  • Achieved a fivefold increase in lycopene production within 3 days.
  • Demonstrated significant improvement over existing metabolic engineering techniques.

Conclusions:

  • MAGE enables rapid, large-scale engineering and evolution of cells.
  • This multiplex approach accelerates the design and evolution of organisms with novel properties.
  • MAGE is a powerful tool for optimizing metabolic pathways for industrial applications.