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SEMPER:通过真核糖核糖体对mRNA多基斯特龙的刻度表达,以实现紧的,可调节比例的多基因表达
Mengtong Duan1, Ishaan Dev2, Andrew Lu3
1Division of Biology and Biological Engineering, Caltech, Pasadena, CA 91125, USA.
Cell systems
|July 6, 2024
概括
我们开发了一种名为SEMPER的新方法,可以从单个mRNA中表达多种蛋白质. 这种技术允许调节的蛋白质比例,使得高效的复合物形成和合成生物学应用中的毒性降低.
科学领域:
- 分子生物学分子生物学
- 合成生物学 合成生物学
- 生物化学 生化学
背景情况:
- 传统的方法往往难以在所需的比例上共同表达多种蛋白质.
- 控制蛋白质固态度对于组装多蛋白质复合体和最大限度地减少细胞负担至关重要.
研究的目的:
- 引入一种新的方法,即通过真核核糖核体 (SEMPER) 来表达mRNA多基斯特龙的固体测量表达,用于可调节的多基斯特龙基因表达.
- 证明SEMPER在各种生物环境中的多功能性,包括蛋白质复合体的形成和抗体的产生.
主要方法:
- 利用翻译启动的漏洞扫描模型来控制单个成绩单上相邻的开放阅读框架 (ORF) 的表达水平.
- 编码光蛋白质,气囊声学记者基因和重组单克隆抗体的工程多晶构造.
- 在哺乳动物细胞系和体外转录中验证了SEMPER.
主要成果:
- 在可调节的比率下,从单个mRNA中成功表达了多达三种光蛋白.
- 使用SEMPER编码的气囊基因,有效形成多蛋白质复合体,细胞毒性降低.
- 从体外转录的mRNA中实现了功能性重组单克隆抗体和光蛋白的多晶表达.
结论:
- SEMPER提供了一种强大而灵活的平台,用于从单个转录物中精确地控制蛋白质表达的固体测量.
- 这种方法对合成生物学,蛋白质工程和治疗性抗体生产有广泛的影响.
- 开发了一个概率模型,以进一步了解SEMPER的潜在机制.
相关概念视频
01:23Regulation of Expression at Multiple Steps
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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...
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01:36What is Gene Expression?
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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
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02:24Regulation of Expression Occurs at Multiple Steps
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Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
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02:33Combinatorial Gene Control
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Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
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02:33Initiation of Translation
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Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...
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02:24Ribosome Profiling
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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
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Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
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