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Related Concept Videos

DNA Topoisomerases02:02

DNA Topoisomerases

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Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
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DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...
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DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied.  After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
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Related Experiment Video

Updated: Feb 10, 2026

Stimulation of Cytoplasmic DNA Sensing Pathways In Vitro and In Vivo
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In vitro DNA SCRaMbLE.

Yi Wu1,2,3, Rui-Ying Zhu1,2, Leslie A Mitchell3

  • 1Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, 300072, Tianjin, China.

Nature Communications
|May 24, 2018
PubMed
Summary

Researchers developed an in vitro SCRaMbLE system for DNA library construction. This method optimizes biosynthetic pathways by rearranging genetic elements, offering a new tool for synthetic biology and genetic engineering.

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Area of Science:

  • Synthetic biology
  • Genetic engineering
  • Molecular biology

Background:

  • Synthetic biology enables heterologous pathway expression and genome-scale synthesis.
  • Designing complex biological networks de novo is challenging.
  • Genetic flexibility is crucial for downstream engineering efforts.

Purpose of the Study:

  • To develop an in vitro DNA library construction method based on structural variation.
  • To enable optimization of biosynthetic pathways through genetic rearrangement.
  • To provide a system for correlating phenotype and genotype.

Main Methods:

  • Developed the in vitro SCRaMbLE system using Cre recombinase and purified DNA with loxPsym sites.
  • Applied top-down and bottom-up in vitro SCRaMbLE to a yeast β-carotene pathway.
  • Demonstrated rearrangement of transcription units to optimize pathway flux.

Main Results:

  • Successfully demonstrated in vitro SCRaMbLE for DNA library construction.
  • Showcased optimization of a β-carotene biosynthetic pathway in yeast.
  • Established a straightforward method for correlating genotype and phenotype.

Conclusions:

  • The in vitro SCRaMbLE system offers a powerful strategy for introducing genetic flexibility.
  • This method facilitates the optimization of biosynthetic pathways via transcription unit rearrangement.
  • The system is amenable to biochemical optimization, surpassing limitations of in vivo systems.