在Cryptococcus中通过RNA干扰使基因沉默
1Department of Biology, Western Carolina University, Cullowhee, NC, USA. ibose@email.wcu.edu.
Methods in molecular biology (Clifton, N.J.)
|May 17, 2024
概括
通过减少mRNA和蛋白质水平,RNA干扰 (RNAi) 会使基因沉默. 这项研究详细介绍了一种双促进子向量方法,通过双链RNA生产在Cryptococcus中有效地静止基因.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
- 生物技术是生物技术.
背景情况:
- RNA干扰 (RNAi) 是一种用于基因沉默的保存生物过程.
- 基因沉默通过向信使RNA (mRNA) 来降解,减少蛋白质的产生来实现.
- 序列特定的双链RNA (dsRNA) 对于激活RNAi通路至关重要.
研究的目的:
- 描述一种使用RNA干扰在Cryptococcus中静止基因的方法.
- 提出一种新型的双促进体载体,用于在体中产生dsRNA.
- 用一个记者基因来评估RNAi系统的效率.
主要方法:
- 开发含有两个融合的GAL7促进体的pIBB103等离子体.
- 在双向转录的促进体之间插入目标基因片段.
- 使用 ura5 片段作为记者来确认RNAi介导的基因沉默.
- 将载体引入Cryptococcus细胞以激活RNAi机制.
主要成果:
- 双促进器系统成功地从插入的目标基因片段生成了dsRNA.
- 双向转录导致RNAi通路的激活.
- ura5 记者片段证实了基因沉默的效率,这种基因沉默是由结构介导的.
结论:
- 描述的双促进子向量系统为Cryptococcus的序列特定基因沉默提供了一个有效的策略.
- 这种方法可以通过减少基因表达来研究基因功能,而不会改变基因组.
- RNAi方法为像Cryptococcus这样的真核生物体的基因操纵提供了一个强大的工具.
相关概念视频
Experimental RNAi
6.1K
RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
6.1K
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...
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
siRNA - Small Interfering RNAs
16.7K
Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...
16.7K
Small interfering RNAs (siRNA)
3.5K
3.5K
MicroRNAs
3.0K
MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
3.0K
Types of RNA
63.6K
Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
63.6K


