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相关概念视频

Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

13.8K
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...
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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.
Types and Mechanism of action
Topoisomerases are divided into two main types. ...
30.7K
DNA Helicases00:55

DNA Helicases

21.0K
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...
21.0K
The DNA Replication Fork01:02

The DNA Replication Fork

35.3K
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...
35.3K
The Replisome03:01

The Replisome

32.8K
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...
32.8K
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

5.7K
DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
5.7K

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相关实验视频

Updated: May 20, 2025

Studying DNA Looping by Single-Molecule FRET
11:27

Studying DNA Looping by Single-Molecule FRET

Published on: June 28, 2014

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通过R-循环中间体通过Cas12a对超级卷轴依赖的DNA查询的动态基础.

Kevin D P Aris1,2, Joshua C Cofsky3,4, Honglue Shi5,6

  • 1Biophysics Program, Stanford University, Stanford, CA, USA.

Nature communications
|March 26, 2025
PubMed
概括
此摘要是机器生成的。

这就是CRISPR-Cas12a.

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Analyzing and Building Nucleic Acid Structures with 3DNA
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Analyzing and Building Nucleic Acid Structures with 3DNA

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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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相关实验视频

Last Updated: May 20, 2025

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科学领域:

  • 分子生物学分子生物学
  • 生物物理学的生物物理.
  • 基因工程是一种基因工程.

背景情况:

  • 由于其可编程的DNA结合和分裂,CRISPR-Cas12a是基因工程中的一个多功能工具.
  • Cas12a作为RNA引导的内核酶起作用,形成一个三链R循环结构,用于DNA位参与.

研究的目的:

  • 研究CRISPR-Cas12a中R循环形成的动态和机制.
  • 通过单分子扭矩光谱来以基对分辨率解决R循环形成.

主要方法:

  • 单分子扭矩光谱学
  • 分析了CRISPR-Cas12a的奥托洛格.
  • 基对分辨率动态研究的基对解析研究.

主要成果:

  • 在R循环形成过程中直接观察动力中间体 (~5bp和~17bp).
  • 在20bp R循环之外识别短暂的DNA解.
  • R-循环形成的景观取决于Cas12a正统,目标序列,不匹配和DNA超级卷.

结论:

  • 一个四态运动模型准确地描述了Cas12a R循环动态.
  • 为了解Cas12a活动和特异性提供了一个生物物理框架.
  • 突出了R循环形成的复杂多态性质.