Applying synthetic biology to the development of in vivo technologies for the monitoring and control of vector-borne diseases.

应用合成生物学来开发用于监测和控制媒介传播疾病的体内技术。

• 批准号: BB/Y008340/1
• 负责人: Tony Nolan
• 金额: $ 133.97万
• 依托单位: Liverpool School of Tropical Medicine
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FY008340%2F1
• 关键词: Applying; synthetic; biology; development; vivo; Applying; synthetic; biology; development; vivo

The ability to genetically modify insects of medical and agricultural importance has opened up the potential for intentionally introducing genetic traits into insect populations. This can be done to alter their ability to reproduce, cause crop damage, or transmit disease-causing pathogens. However, there are challenges in getting the introduced traits to spread throughout the population. Usually, the added genetic trait does not improve the fitness of the insects carrying it, and sometimes it even has a negative effect. This means that a large number of modified insects, often tens of millions, need to be released to have a significant impact on the population. This process is costly and logistically challenging, and the effects only last as long as the continuous release of large numbers of modified insects.Recent advancements in genetic control, such as "gene drive," address this problem by biasing the inheritance of the modification in each generation. This means that the frequency of the modified trait can increase rapidly in the population. These approaches show promise because they are self-sustaining, requiring only a few released insects to have a long-term effect, and they are species-specific since the modified traits are passed on through mating between insects of the same species. Many gene drive designs utilize genome editing tools like CRISPR to bias the inheritance of the gene drive element in sperm or eggs, which are then passed on to the next generation. By making small changes to the CRISPR element, such as its duration and timing of DNA cleavage activity, its performance in terms of inheritance can be drastically affected. Limiting the expression of the gene drive element to the germline cells, where it needs to be active, can greatly enhance the fitness of insects carrying the gene drive and increase its chances of spreading through the target population. Additionally, many gene drives contain a genetic "cargo" that produces a desired effect in the insects carrying it, such as activating the immune system against a pathogen or interfering with parasite replication. Expressing these effects only in non-infected insects or specific tissues can be costly. Having the ability to fine-tune the expression of the gene drive and its cargo within the insect, both in terms of timing and location, can significantly improve their effectiveness by effectively minimising expression of the different elements to only the cells or conditions where it is essential that they act.In this proposal, we aim to:1. Complete the process of describing gene expression at the single-cell level in the ovary and testis. We will extract information about the DNA sequences of the genetic switches that control expression in the relevant cells that can ensure biased inheritance of the gene drive with minimal or no unwanted effects. We will design a cell-based approach to test different control sequences efficiently and then evaluate the most promising combinations in laboratory populations of modified mosquitoes.2. Enhance the specificity of gene drive expression and/or its cargo by making sure they are only active in response to specific signals, such as specific RNA sequences from the pathogen. These RNA-based "riboswitches" are innovative, and demonstrating their effectiveness in this system would have wide-ranging implications, not just in insect control but also in improving the precision and specificity of genome editing in various applications, including healthcare applications like in vivo genome editing and CRISPR-based diagnostic assays.

遗传修改医学和农业重要性昆虫的能力为将遗传特征引入昆虫种群的可能性打开了潜力。这可以改变其繁殖，造成农作物损害或传播引起疾病的病原体的能力。但是，要使引入的特征传播到整个人群中存在挑战。通常，增加的遗传特征不会改善携带昆虫的适应性，有时甚至具有负面影响。这意味着需要释放大量改良的昆虫，通常需要数千万，以对人口产生重大影响。这个过程在逻辑上是昂贵和具有挑战性的，只要连续释放大量修改昆虫。这意味着在人群中，修改性状的频率可以迅速增加。这些方法表现出希望，因为它们是自我维持的，只需要少数释放的昆虫才能长期作用，并且它们是特定于物种的，因为修饰的性状通过同一物种的昆虫之间的交配传递。许多基因驱动器设计都使用CRISPR等基因组编辑工具来偏向精子或卵中基因驱动元件的遗传，然后将其传递给下一代。通过对CRISPR元素进行小改变，例如其持续时间和DNA裂解活动的时机，其在遗传方面的性能可能会受到巨大影响。将基因驱动元件的表达限制在需要活跃的种系细胞上，可以极大地提高携带基因驱动的昆虫的适应性，并增加其在目标群体中传播的机会。此外，许多基因驱动器都包含遗传“货物”，该遗传“货物”在携带昆虫的昆虫中产生所需的作用，例如激活免疫系统针对病原体或干扰寄生虫复制。仅在未感染的昆虫或特定组织中表达这些作用的代价很高。在时间和位置方面，具有微调基因驱动器及其在昆虫中的货物的表达能力，可以通过有效地最大程度地减少不同元素表达的表达来显着提高其有效性，仅对它们在此提案中至关重要的细胞或条件。我们的目标是：1。完成卵巢和睾丸中单细胞水平上描述基因表达的过程。我们将提取有关控制在相关细胞中表达表达的遗传开关的DNA序列的信息，以确保具有最小或无需效应的基因驱动的偏置遗传。我们将设计一种基于细胞的方法来有效测试不同的控制序列，然后评估改良蚊子实验室种群中最有希望的组合。2。通过确保仅对特定信号（例如病原体的特定RNA序列）进行活性来增强基因驱动表达和/或其货物的特异性。这些基于RNA的“核糖开关”具有创新性，并且证明它们在该系统中的有效性不仅在昆虫控制方面具有广泛的含义，而且还可以提高基因组编辑在各种应用中的精度和特异性，包括医疗保健应用，包括INTEMAIME基因组编辑和基于CRISPR的诊断诊断。