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相关概念视频

Viral Mutations00:36

Viral Mutations

32.1K
A mutation is a change in the sequence of bases of DNA or RNA in a genome. Some mutations occur during replication of the genome due to errors made by the polymerase enzymes that replicate DNA or RNA. Unlike DNA polymerase, RNA polymerase is prone to errors because it is not capable of “proofreading” its work. Viruses with RNA-based genomes, like HIV, therefore accrue mutations faster than viruses with DNA-based genomes. Because mutation and recombination provide the raw material...
32.1K
Leaky Scanning02:28

Leaky Scanning

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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.1K
Mismatch Repair01:20

Mismatch Repair

4.8K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
4.8K
Proofreading01:43

Proofreading

53.7K
Overview
53.7K
RNA Editing02:23

RNA Editing

8.9K
RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
8.9K
Improving Translational Accuracy02:07

Improving Translational Accuracy

8.6K
Base complementarity between the three base pairs of mRNA codon and the tRNA anticodon is not a failsafe mechanism. Inaccuracies can range from a single mismatch to no correct base pairing at all. The free energy difference between the correct and nearly correct base pairs can be as small as 3 kcal/ mol. With complementarity being the only proofreading step, the estimated error frequency would be one wrong amino acid in every 100 amino acids incorporated. However, error frequencies observed in...
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相关实验视频

Updated: Jun 3, 2025

Reverse Genetics to Engineer Positive-Sense RNA Virus Variants
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Reverse Genetics to Engineer Positive-Sense RNA Virus Variants

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通过进化逆转录酶促进对伪尿素-CMC的读透和突变.

Zhiyong He1, Weiqi Qiu1, Huiqing Zhou2

  • 1Department of Chemistry, Merkert Chemistry Center, Boston College, Chestnut Hill, MA, USA.

Communications biology
|January 11, 2025
PubMed
概括

这项研究介绍了Mut-Ψ-seq,这是一种用于精确地绘制RNA中的伪尿素 (Ψ) 的新方法,即使是在具有挑战性的富含U的区域. 它克服了现有技术的局限性,增强了表观转录学研究.

科学领域:

  • 史诗转录组学 史诗转录组学
  • 基因组RNA的修改 基因组RNA的改变
  • 分子生物学分子生物学

背景情况:

  • 伪乌里丁 (Ψ) 是一种普遍存在的RNA修饰,对生物功能至关重要.
  • 现有的方法很难在基础分辨率上检测 Ψ,特别是在 Ψ 常见的 U 丰富序列中.

研究的目的:

  • 开发一种新的方法来绘制伪尿素 (Ψ) 的基本分辨率映射.
  • 克服在富含U的RNA序列中检测 Ψ 的局限性.
  • 扩大用于表观转录学研究的工具包.

主要方法:

  • 开发Mut-Ψ-seq,结合N-环基N'-(2-摩尔福利诺乙基) 碳二胺 (CMC) 化学标记和一个工程反转录酶 (RT-1306).
  • CMC选择性地标记 Ψ 位点,形成一个CMC-Ψ adduct.
  • RT-1306在CMC-Ψ位点表现出增强的读透和突变,使检测成为可能.

主要成果:

  • 在HEK-293T细胞中,使用正交化学处理 (CMC和双硫酸盐) 识别了高可信度伪尿素 (Ψ) 位点,这些位点在HEK-293T细胞中富含多A的RNA中.
  • 突变特征在含有UU的序列中准确地解析了 Ψ 位置,揭示了它们的多样性.
  • 该方法成功地在基本分辨率上映了 Ψ,包括在具有挑战性的 U 丰富的环境中.

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Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV
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Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV

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Last Updated: Jun 3, 2025

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Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells
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Determining 3'-Termini and Sequences of Nascent Single-Stranded Viral DNA Molecules during HIV-1 Reverse Transcription in Infected Cells

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Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV
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Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV

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结论:

  • Mut-Ψ-seq为基本分辨率伪尿素 (Ψ) 映射提供了一个强大的方法.
  • 该技术增强了研究 Ψ 在 U 丰富序列中的能力,这对表皮转录学来说是一个重大进步.
  • 这项工作为伪尿素 (Ψ) 研究和表体转录学研究提供了有价值的方法和数据集.