Jove
Visualize
联系我们
JoVE
x logofacebook logolinkedin logoyoutube logo
关于 JoVE
概览领导团队博客JoVE 帮助中心
作者
出版流程编辑委员会范围与政策同行评审常见问题投稿
图书馆员
用户评价订阅访问资源图书馆顾问委员会常见问题
研究
JoVE JournalMethods CollectionsJoVE Encyclopedia of Experiments存档
教育
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab Manual教师资源中心教师网站
使用条款与条件
隐私政策
政策

相关概念视频

RNA Splicing01:32

RNA Splicing

60.3K
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
60.3K
Alternative RNA Splicing02:18

Alternative RNA Splicing

24.7K
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
24.7K
Alternative RNA Splicing02:18

Alternative RNA Splicing

4.8K
4.8K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.6K
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...
6.6K
Pre-mRNA Processing: RNA Splicing01:36

Pre-mRNA Processing: RNA Splicing

6.6K
6.6K
RACE - Rapid Amplification of cDNA Ends02:35

RACE - Rapid Amplification of cDNA Ends

7.1K
Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...
7.1K

您也可能阅读

相关文章

通过共同作者、期刊和引用图与本文相关的文章。

排序
Same author

The BmCPV-derived viral small peptide vsp1S4(-) suppresses viral replication by triggering apoptosis via the ROS-JNK signalling pathway.

Cellular and molecular life sciences : CMLS·2026
Same author

Human MAITregs possess diverse TCR repertoires and are functionally supported by glycolysis.

Science China. Life sciences·2026
Same author

Impact of cricothyroid membrane puncture anesthesia on elderly patients undergoing endotracheal intubation after anesthesia induction: a clinical study.

Frontiers in physiology·2026
Same author

Genome-scale DNA methylome and transcriptome profiling of midgut of Bombyx mori infected with BmCPV.

Scientific data·2026
Same author

Energy production is suppressed by vcircRNA_000048- bom-miR-2753-METTL20 regulatory axis in the BmCPV-infected silkworm, Bombyx mori.

Insect biochemistry and molecular biology·2026
Same author

Pulmonary nodules with explosive calcification were finally diagnosed as lung adenocarcinom: A case report.

Medicine·2026

相关实验视频

Updated: Jan 15, 2026

Use of Alu Element Containing Minigenes to Analyze Circular RNAs
13:10

Use of Alu Element Containing Minigenes to Analyze Circular RNAs

Published on: March 10, 2020

7.7K

为增强线性RNA表达增强背接类似的工程站点.

Xialing Chen1, Sufei Sun1, Kaimeng Li2

  • 1School of Life Sciences, Suzhou Medical College of Soochow University, Suzhou, 215123, China.

Applied microbiology and biotechnology
|October 16, 2025
PubMed
概括

将外源基因引入细胞可以形成圆形RNA (circRNA) 或线性RNA. 修改这些基因的背部拼接样 (BSL) 位点可以增强基因的表达,从而提供一种新的基因表达系统.

关键词:
卢克基因 卢克基因来自Luc的环RNAs.一个类似背接的网站.

更多相关视频

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
10:06

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

9.4K
A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
08:53

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

3.2K

相关实验视频

Last Updated: Jan 15, 2026

Use of Alu Element Containing Minigenes to Analyze Circular RNAs
13:10

Use of Alu Element Containing Minigenes to Analyze Circular RNAs

Published on: March 10, 2020

7.7K
Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
10:06

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

9.4K
A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
08:53

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

3.2K

科学领域:

  • 分子生物学分子生物学
  • 在RNA生物学,RNA生物学.
  • 基因表达 基因表达

背景情况:

  • 循环RNA (circRNA) 和从前RNA形成线性RNA是一个由RNA序列,调节元件和RNA结合蛋白影响的竞争过程.
  • 培养细胞中的外源基因表达涉及复杂的拼接机制.

研究的目的:

  • 研究不同细胞系中的外源性光酶 (Luc) 基因形成circRNA的过程.
  • 分析背部拼接样 (BSL) 位点在Luc衍生的circRNAs的形成和表达中的作用.
  • 探索调节BSL遗址的潜力,以开发高效的基因表达系统.

主要方法:

  • 在HepG2,BmN和CIK细胞中引入了露西法酶 (Luc) 表达载体和体外转录的LucRNA.
  • 使用逆转录聚合酶链反应 (RTPCR),桑格测序和逆转录滚动圆放大.
  • 对BSL位点的突变分析及其对RNA表达和光酶活性影响的评估.
  • 抑制与circRNA形成相关的蛋白质基因.

主要成果:

  • 在多个细胞系中成功形成并检测到Luc衍生的circRNAs.
  • 在Luc-circRNA中确定了BSL位点,与正规拼接位点不同.
  • 在BSL位点的突变显著降低了circRNA表达,同时增加了线性Luc mRNA水平和光酶活性.
  • 具有BSL位点突变的重组baculoviruses显示 circRNA形成减少和RNA表达增加.
  • 沉默参与circRNA形成的关键蛋白质基因降低了circRNA效率并增加了Luc表达.

结论:

  • 引入培养细胞的外源基因可以经历拼接,形成circRNAs.
  • 修改外源基因中的BSL位点可以增强它们的表达,提供一种新的调节机制.
  • 对BSL遗址的监管为开发高效和有针对性的基因表达系统提供了一个有希望的战略.