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Genome Size and the Evolution of New Genes03:21

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While every living organism has a genome of some kind (be it RNA, or DNA), there is considerable variation in the sizes of these blueprints. One major factor that impacts genome size is whether the organism is prokaryotic or eukaryotic. In prokaryotes, the genome contains little to no non-coding sequence, such that genes are tightly clustered in groups or operons sequentially along the chromosome. Conversely, the genes in eukaryotes are punctuated by long stretches of non-coding sequence.
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Following the Dynamics of Structural Variants in Experimentally Evolved Populations
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A DNA Inversion System in Eukaryotes Established via Laboratory Evolution.

Peiyan Han1,2, Yuan Ma1,2, Zongheng Fu1,2

  • 1Frontier Science Center for Synthetic Biology (Ministry of Education), Tianjin University, Tianjin, 300072, China.

ACS Synthetic Biology
|August 23, 2021
PubMed
Summary

Scientists engineered a bacterial DNA inversion system for use in eukaryotes. This new system, Rci8 recombinase, offers high specificity for DNA inversions, enabling precise genetic control in yeast and mammalian cells.

Keywords:
directed evolutioneukaryotessite-specific DNA inversionsynthetic biology

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Genetics

Background:

  • Site-specific recombination systems, like DNA inversion, are crucial for bacterial genetic diversity and adaptation through programmed rearrangements.
  • Eukaryotic systems currently lack a DNA inversion tool with a strong directionality bias for precise genetic manipulation.

Purpose of the Study:

  • To develop a highly directional DNA inversion system for eukaryotic applications.
  • To engineer a bacterial tyrosine recombinase for enhanced performance in yeast and mammalian cells.
  • To establish a novel tool for genetic circuits, cellular barcoding, and synthetic genome engineering.

Main Methods:

  • Directed evolution was used to modify the bacterial Rci recombinase.
  • The engineered Rci8 recombinase and specific sfxa101 sites were utilized to create the DNA inversion system.
  • The system's efficiency and specificity were tested in yeast and mammalian cells, including on linear chromosomes.

Main Results:

  • A mutant Rci8 recombinase was identified with a high inversion-to-deletion ratio (up to ~4320) in yeast.
  • The developed system demonstrated specificity for DNA inversions over deletions between directly repeated sites.
  • The reversible DNA inversion system successfully functioned as an on/off transcriptional switch and on linear chromosomes.

Conclusions:

  • A novel, highly directional DNA inversion system has been successfully established in eukaryotes (yeast and mammalian cells).
  • This engineered system provides a powerful new tool for precise genetic engineering applications.
  • The eukaryotic DNA inversion system holds significant potential for advancing genetic circuits, cellular barcoding, and synthetic genomics.