一个基于RNAi和抑制蛋白的调节性遗传开关,用于调节哺乳动物细胞中的基因表达
Tara L Deans1, Charles R Cantor, James J Collins
1Department of Biomedical Engineering, Center for BioDynamics and Center for Advanced Biotechnology, Boston University, Boston, MA 02215, USA.
Cell
|July 31, 2007
概括
研究人员开发了一种可调节的遗传开关,以精确控制哺乳动物细胞中的基因表达. 该系统实现了99%以上的基因沉默,并允许微调基因活动,从而开辟了新的研究途径.
科学领域:
- 分子生物学分子生物学
- 基因规则 基因规则
- 合成生物学 合成生物学
背景情况:
- 精确控制基因表达对于理解基因功能和开发新疗法至关重要.
- 现有的基因沉默方法往往缺乏可调性或完全抑制.
研究的目的:
- 为了设计一种新的基因开关,在哺乳动物细胞中实现强大且可调节的基因沉默.
- 为了证明该系统在调节各种基因的有效性,包括记者基因,毒素和诱导亡的基因.
主要方法:
- 设计了一种工程遗传开关,将抑制蛋白与RNA干扰 (RNAi) 目标结合起来.
- 该开关调节基因表达的能力在小鼠和人类细胞系中使用增强绿色光蛋白 (EGFP) 进行了测试.
- 该系统的模块化和沉默能力通过控制喉毒素和Cre recombinase的表达得到了进一步的验证.
主要成果:
- 基因开关实现了99%以上的基因表达抑制.
- 成功证明了对基因表达水平的可调控控制.
- 该系统有效地调节了喉毒素,Cre复合酶和一个亲亡基因的表达,揭示了诱导亡的值反应.
结论:
- 这种工程基因开关为哺乳动物系统中基因表达的紧密和可调节的控制提供了一个强大的工具.
- 该系统有助于对基因功能进行调查,并识别基因相关表型中的值反应.
- 这项技术对基础研究和需要精确基因调节的治疗应用都有潜力.
相关概念视频
Types of RNA
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...
Riboswitches
Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
Types of 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 regulating 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 Performs Diverse...
RNA Performs Diverse...
Regulation of Expression at Multiple Steps
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
Repressible Operon: trp Operon
The trp operon in Escherichia coli exemplifies a repressible operon. It regulates the synthesis of tryptophan through repressor-mediated transcriptional control and attenuation. This dual regulatory mechanism ensures tryptophan biosynthesis occurs only when needed, conserving cellular resources.Structure of the trp OperonThe trp operon consists of five structural genes (trpE, trpD, trpC, trpB, and trpA) that encode enzymes for tryptophan biosynthesis. These genes are transcribed as a single...
Transcriptional Regulation: Riboswitches
Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...


