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Related Concept Videos

Single-Strand DNA Binding Proteins01:03

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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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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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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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The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
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Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
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Related Experiment Video

Updated: Jun 4, 2025

Substrate Generation for Endonucleases of CRISPR/Cas Systems
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Cas12e orthologs evolve variable structural elements to facilitate dsDNA cleavage.

Danyuan Li1,2, Shouyue Zhang3,4, Shuo Lin1,2

  • 1Beijing Frontier Research Center for Biological Structure, State Key Laboratory of Membrane Biology, School of Life Sciences, Tsinghua University, Beijing, 100084, China.

Nature Communications
|December 31, 2024
PubMed
Summary
This summary is machine-generated.

Researchers explored diverse CRISPR-Cas systems, specifically Cas12e nucleases, for DNA manipulation. They discovered new Cas12e variants with unique properties, enhancing genome editing tools.

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Last Updated: Jun 4, 2025

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Type V CRISPR-Cas systems, including Cas12e nucleases, are versatile tools for DNA manipulation and genome editing.
  • Existing Cas12e variants like DpbCas12e and PlmCas12e show distinct in vitro DNA cleavage activities, highlighting family diversity.

Purpose of the Study:

  • To comprehensively characterize the Cas12e family by identifying and analyzing novel members.
  • To understand the structural basis for the diverse enzymatic properties and cleavage efficacies within the Cas12e family.

Main Methods:

  • Identification and characterization of six new Cas12e members.
  • Analysis of CRISPR-locus architectures, PAM preferences, and in vitro dsDNA cleavage activities.
  • Structural comparisons of Cas12e variants, focusing on the NTSB domain and its role in DNA unwinding.

Main Results:

  • Six previously unreported Cas12e members were identified with varying CRISPR-locus architectures, PAM preferences, and cleavage efficiencies.
  • PlmCas12e demonstrated superior trans-cleavage activity and lower salt sensitivity in cis-cleavage compared to other variants.
  • Structural analysis revealed the NTSB domain's importance for DNA unwinding at high salt concentrations, with some variants utilizing charged loops instead.

Conclusions:

  • Divergent evolution of structural elements, such as the NTSB domain, drives nuclease diversity within the Cas12e family.
  • These structural variations contribute to the adaptation of Cas12e nucleases to different environmental conditions and enhance their utility as genome editing tools.