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Ribozymes02:47

Ribozymes

12.3K
The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can...
12.3K
Restriction Enzymes01:11

Restriction Enzymes

30.7K
Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...
30.7K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.0K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
6.0K
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

11.2K
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
11.2K
RNA Interference01:23

RNA Interference

26.0K
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
26.0K
Homologous Recombination02:31

Homologous Recombination

50.5K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
50.5K

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Related Experiment Video

Updated: Jul 4, 2025

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
09:12

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation

Published on: September 16, 2019

8.3K

Hydrolytic endonucleolytic ribozyme (HYER) is programmable for sequence-specific DNA cleavage.

Zi-Xian Liu1, Shouyue Zhang1, Han-Zhou Zhu1

  • 1Beijing Advanced Innovation Center for Structural Biology, State Key Laboratory of Membrane Biology, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.

Science (New York, N.Y.)
|February 1, 2024
PubMed
Summary
This summary is machine-generated.

Researchers discovered HYdrolytic Endonucleolytic Ribozymes (HYERs), a type of catalytic RNA, that can cut DNA. These natural ribozymes show potential for precise DNA manipulation and genome editing applications.

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Last Updated: Jul 4, 2025

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Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Ribozymes are catalytic RNA molecules with essential roles in cellular processes.
  • Natural ribozymes are increasingly recognized for their potential beyond RNA catalysis, including DNA manipulation.

Purpose of the Study:

  • To identify and characterize natural ribozymes functioning as sequence-specific DNA endonucleases.
  • To explore the repurposing of these ribozymes as novel DNA manipulation tools.

Main Methods:

  • Focusing on bacterial group II-C introns, researchers screened for systems lacking intron-encoded proteins.
  • In vitro assays were used to test the cleavage activity of identified ribozymes (HYERs) against various nucleic acid substrates.
  • Cryo-electron microscopy was employed to determine the structural basis of HYER1's DNA binding and catalytic mechanism.
  • Rational design strategies were applied to engineer HYER variants with enhanced specificity and functionality.

Main Results:

  • Several intron systems, termed HYdrolytic Endonucleolytic Ribozymes (HYERs), were identified that cleave RNA, single-stranded DNA, and double-stranded DNA (dsDNA).
  • HYER1 demonstrated the ability to induce dsDNA breaks in mammalian genomes in vitro.
  • Cryo-electron microscopy revealed a homodimeric structure of HYER1 with a Mg2+-dependent hydrolysis pocket and DNA-binding capability.
  • Engineered HYER variants exhibited improved DNA manipulation specificity and flexibility.

Conclusions:

  • Natural ribozymes, specifically HYERs from group II-C introns, possess DNA endonuclease activity.
  • These HYERs can be engineered for precise DNA cleavage, offering a promising new platform for genome editing and biotechnology.