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相关概念视频

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...
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Restriction Enzymes01:11

Restriction Enzymes

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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...
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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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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...
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Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

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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...
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RNA Interference01:23

RNA Interference

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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...
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Homologous Recombination02:31

Homologous Recombination

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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...
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相关实验视频

Updated: Jul 4, 2025

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

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水解内核解酶 (HYER) 可编程用于特定序列的DNA裂变

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
概括

研究人员发现了HYERs,一种可以切割DNA的催化RNA. 这些天然的 ribozymes 显示出精确的 DNA 操纵和基因组编辑应用的潜力.

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Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
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Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

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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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相关实验视频

Last Updated: Jul 4, 2025

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

Published on: September 16, 2019

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Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
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Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

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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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科学领域:

  • 分子生物学
  • 生物化学
  • 遗传学

背景情况:

  • ribozymes 是催化性RNA分子,在细胞过程中起着重要的作用.
  • 自然的 ribozymes 越来越多地被认为具有超越 RNA 催化,包括 DNA 操纵的潜力.

研究的目的:

  • 识别和描述作为序列特异性DNA内核酶的自然 ribozymes.
  • 探索这些 ribozymes 作为新型的 DNA 操纵工具.

主要方法:

  • 研究人员专注于细菌II-C组内蛋白, 选缺乏内蛋白编码的系统.
  • 在体外测试中测试了已识别的核酶 (HYER) 与各种核酸基质的分裂活性.
  • 使用冷电子显微镜确定HYER1的DNA结合和催化机制的结构基础.
  • 应用了合理的设计策略来设计具有增强特异性和功能的HYER变体.

主要成果:

  • 鉴定出了几种称为HYER的内离子系统,可以分裂RNA,单链DNA和双链DNA.
  • 在实验室中,HYER1显示能够诱导哺乳动物基因组中的dDNA断裂.
  • 低温电子显微镜显示HYER1具有Mg2+依赖的水解口袋和DNA结合的能力.
  • 改造的HYER变种表现出更好的DNA操纵特异性和灵活性.

结论:

  • 自然的核酶,特别是来自II-C组内核的HYER,具有DNA内核酶活性.
  • 这些HYER可以被设计为精确的DNA分裂,为基因组编辑和生物技术提供了一个有前途的新平台.