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Related Concept Videos

The Replisome03:01

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
The DNA Replication Fork01:02

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...
Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
Lagging Strand Synthesis01:59

Lagging Strand Synthesis

During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
Replication in Prokaryotes01:32

Replication in Prokaryotes

DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the single origin of replication (ori) on the cell's circular chromosome. Replication then proceeds around the entire circle of the chromosome in each direction from the two replication forks, resulting in two DNA molecules.
Many Proteins Work Together to Replicate the Chromosome
Replication is coordinated and carried out by a host of specialized...
Replication in Prokaryotes02:35

Replication in Prokaryotes

Overview

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Folding and Characterization of a Bio-responsive Robot from DNA Origami
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Published on: December 3, 2015

Bioproduction of ∼10 knt single-stranded DNA for constructing large DNA origami structures.

Meiling Lu1,2, Xiwei Wang2, Baohong He2

  • 1State Key Laboratory of Discovery and Utilization of Functional Components in Traditional Chinese Medicine & School of Pharmaceutical Sciences, Guizhou Medical University, Guiyang, 561113, China.

Materials Today. Bio
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Summary

Researchers developed a new method to produce long single-stranded DNA (ssDNA) for creating larger DNA origami nanostructures. This advancement enables more functional sites and scalable production of complex DNA-based nanomaterials.

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Production of Dynein and Kinesin Motor Ensembles on DNA Origami Nanostructures for Single Molecule Observation
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Production of Dynein and Kinesin Motor Ensembles on DNA Origami Nanostructures for Single Molecule Observation

Published on: October 15, 2019

Area of Science:

  • Nanotechnology
  • Molecular Biology
  • Biotechnology

Background:

  • Traditional DNA origami structures using M13 scaffolds have limitations in size and functional integration.
  • Scaling up DNA origami requires longer single-stranded DNA (ssDNA) scaffolds, presenting challenges in preparation and stable synthesis.

Purpose of the Study:

  • To establish an efficient biosynthesis platform for producing long ssDNA (approximately 10 kilobase pairs).
  • To utilize the synthesized long ssDNA for constructing large, stable DNA origami nanostructures with high yield.
  • To overcome the limitations of existing DNA origami scaffolds for enhanced functionalization and size expansion.

Main Methods:

  • Developed a phage-phagemid production system for long ssDNA biosynthesis.
  • Optimized synthesis parameters in shake flasks and bioreactors to enhance ssDNA yield.
  • Assembled large triangular and rectangular DNA origami structures using the produced long ssDNA scaffolds.

Main Results:

  • Achieved an approximately tenfold increase in long ssDNA yield through optimized biosynthesis.
  • Successfully constructed large DNA origami structures (e.g., 161 nm triangular, 93 × 115 nm rectangular).
  • The new structures offer approximately 100 additional potential functionalization sites compared to M13mp18-based structures.

Conclusions:

  • The study presents a scalable method for producing long ssDNA scaffolds, crucial for advanced DNA nanotechnology.
  • The developed platform provides a practical foundation for fabricating large DNA origami nanostructures with enhanced functionality.
  • This work facilitates the creation of more complex and versatile DNA-based nanomaterials.