绿门2.0:向后兼容的附加组件,用于组装复杂的转录单元,并将其与绿门堆叠
Marcel Piepers1, Katarina Erbstein1, Jazmin Reyes-Hernandez1
1Center for Organismal Studies (COS), University of Heidelberg, Heidelberg, Germany.
PloS one
|September 8, 2023
概括
绿门2.0增强了用于植物合成生物学的模块化克隆. 这种新系统允许更复杂的转录单元和高效的多基因组装,改善基因工程工作流程.
科学领域:
- 分子生物学分子生物学
- 合成生物学 合成生物学
- 植物生物技术 植物生物技术
背景情况:
- 像GreenGate这样的模块化克隆系统对于植物基因工程至关重要,它可以从标准化构件组装转录单元 (TU).
- 现有的系统虽然很受欢迎,但在模块容量,组装效率和简化入门克隆生成方面存在局限性.
- 这些局限性阻碍了复杂的应用,如多重基因组编辑和大规模的多基因结构组装.
研究的目的:
- 推出GreenGate 2.0 (GG2.0),这是一个先进的模块化克隆系统,旨在克服原始GreenGate系统的限制.
- 在植物研究中提高组装复杂的转录单元和多基因结构的多功能性和效率.
- 为CRISPR/Cas9基因组编辑和诱导基因表达等先进应用提供新的工具和模块.
主要方法:
- 开发GG2.0具有扩展的悬架,以容纳1级TU中的最多12个0级模块.
- 引入通用输入生成器等离子体 (pUEG) 以简化0级模块的创建.
- 实施GreenBraid,一种用于多基因组件的TUs代堆叠的新方法.
主要成果:
- GG2.0允许从最多12个模块组装1级TU,显著增加复杂性.
- 格林布雷德方法为代多基因组装提供了一种高效和方便的方法.
- GG2.0包括用于CRISPR/Cas9多重基因组编辑的专门模块,以及用于可诱导基因表达和光标记物的预组装载体.
- 该系统与大多数现有的GreenGate模块保持了向后兼容性.
结论:
- 绿门2.0显著增强了植物合成生物学模块化克隆的能力.
- 扩大的模块容量和高效的多基因组装方法简化了复杂遗传结构的创建.
- GG2.0提供了多功能工具,将加速植物基因组编辑和基因表达研究等领域的研究.
相关概念视频
Cooperative Binding of Transcription Regulators
6.5K
Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome. Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form...
6.5K
Assembly of Signaling Complexes
5.8K
Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
5.8K
Protein Complex Assembly
2.1K
2.1K
Master Transcription Regulators
6.9K
Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
6.9K
Combinatorial Gene Control
8.4K
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...
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...
8.4K
Eukaryotic Transcription Activators
11.1K
Transcription activators are proteins that promote the transcription of genes from DNA to RNA. In most cases, these proteins contain two separate domains ‒ a domain that binds to DNA and a domain for activating transcription; however, in some cases, a single domain is responsible for both binding and activation of transcription, as seen in the glucocorticoid receptor and MyoD.
The binding domains are capable of recognizing and interacting with regulatory sequences on the DNA. These...
The binding domains are capable of recognizing and interacting with regulatory sequences on the DNA. These...
11.1K


