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Related Concept Videos

The Evidence for Evolution02:55

The Evidence for Evolution

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Genetic variations accumulating within populations over generations give rise to biological evolution. Evolutionary changes can result in the formation of novel varieties and entire new species. These changes are responsible for the diverse forms of life inhabiting the planet. The evidence for evolution suggests that all living organisms descended from common ancestors.
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Convergent Evolution01:54

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Evolution shapes the features of organisms over time, ensuring that they are suited for the environments in which they live. Sometimes, selection pressure leads to the rise of similar but unrelated adaptations in organisms with no recent common ancestors, a process known as convergent evolution.
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Natural selection is an evolutionary process in which individuals with survival-promoting traits reproduce at higher rates. These favorable traits become more common within a population or species. Naturally selected traits initially arise via random genetic mutations. In order for selection to occur, there must be variation within a population, the trait controlling the variation must be heritable, and there must be an evolutionary advantage for variation in the trait.
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Somatic to iPS Cell Reprogramming01:29

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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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Related Experiment Video

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Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
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Directed Evolution of Reprogramming Factors by Cell Selection and Sequencing.

Veeramohan Veerapandian1, Jan Ole Ackermann2, Yogesh Srivastava1

  • 1CAS Key Laboratory of Regenerative Biology, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, and Guangzhou Medical University, Guangzhou, China; Genome Regulation Laboratory, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China; University of Chinese Academy of Sciences, No.19A Yuquanlu, Beijing, China.

Stem Cell Reports
|August 7, 2018
PubMed
Summary

We developed DERBY-seq, a novel method for evolving reprogramming factors. This technique successfully enhanced transcription factors like SOX2 and converted SOX17 into potent pluripotency inducers.

Keywords:
OCT4SOX17SOX2cellular reprogrammingdeep mutational scanningdirected evolutionprotein engineeringsynthetic biologysynthetic transcription factors

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Area of Science:

  • Cell biology
  • Molecular biology
  • Biotechnology

Background:

  • Directed biomolecular evolution enhances proteins but hasn't been applied to mammalian cell reprogramming.
  • Identifying effective reprogramming factors is crucial for cell fate control.

Purpose of the Study:

  • To introduce and validate DERBY-seq (directed evolution of reprogramming factors by cell selection and sequencing) for discovering enhanced reprogramming transcription factors.
  • To demonstrate the method's efficacy using SOX2 and SOX17 in pluripotency reprogramming.

Main Methods:

  • Pooled screening of positionally randomized gene libraries.
  • Cell selection based on phenotypic readouts.
  • Genotyping by amplicon sequencing for candidate identification.

Main Results:

  • Identified several SOX2 variants that outperform wild-type SOX2 in reprogramming cocktails.
  • Discovered highly effective SOX17 variants that induce pluripotency, demonstrating SOX17's convertibility.
  • Validated DERBY-seq's capability to evolve reprogramming factors.

Conclusions:

  • DERBY-seq is a powerful, broadly applicable approach for discovering novel reprogramming factors.
  • The method facilitates the engineering of transcription factors for specific cell reprogramming applications.
  • DERBY-seq holds promise for advancing direct lineage reprogramming in vitro and in vivo.