21-BBSRC/NSF-BIO: Developing large serine integrases as tools for constructing and manipulating synthetic replicons.

21-BBSRC/NSF-BIO：开发大型丝氨酸整合酶作为构建和操作合成复制子的工具。

• 批准号: BB/X012085/1
• 负责人: Femi Olorunniji
• 金额: $ 88.36万
• 依托单位: Liverpool John Moores University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX012085%2F1
• 关键词: 21; BBSRC; NSF; BIO; Developing

This collaborative work develops an expanded genetic toolkit to facilitate asking fundamental questions about how assemblies of genes work together to perform natural or synthetic functions. This toolkit is intended to be flexible and useful for the broad variety of creative genetic manipulations envisioned (past and future) by biologists, including the modular construction and convenient editing of programmable genetic circuits. The core of this toolkit is a family of bacteriophage enzymes termed "Large serine integrases (LSIs)" that mediate precise DNA rearrangements. These rearrangements can be reversed by the addition of a second protein called a recombination directionality factor (RDF). However, RDFs have been difficult to identify in natural sources, and how they work is not well understood. This work uses new approaches to address both of those problems. This project will significantly expand and innovate the LSI-RDF toolkit that is available for use in synthetic cell studies. Our work will provide: a large panel of characterized, orthogonal LSI-RDF pairs; new approaches for future researchers to identify and/or engineer LSI-RDF systems; structural information and detailed understanding needed to drive further engineering, and new light-controlled functionality.The PIs are developing ways to introduce high school students to the excitement of seeing and playing with the 3D structures of DNA, working with community groups to discuss the potential power of modern genetic tools and to encourage enrolment to university STEM-related subjects especially among women and underrepresented groups, and interfacing with the biotechnology industry to improve curriculum for industry-bound students.

这项合作开发了一个扩展的遗传工具包，以方便提出有关基因组装如何协同工作以执行天然或合成功能的基本问题。该工具包旨在灵活且有用地用于生物学家设想（过去和未来）的各种创造性基因操作，包括可编程基因电路的模块化构造和方便编辑。该工具包的核心是称为“大丝氨酸整合酶 (LSI)”的噬菌体酶家族，可介导精确的 DNA 重排。这些重排可以通过添加第二种蛋白质（称为重组方向性因子（RDF））来逆转。然而，天然来源中的 RDF 很难识别，而且它们的工作原理也不清楚。这项工作使用新方法来解决这两个问题。该项目将显着扩展和创新可用于合成细胞研究的 LSI-RDF 工具包。我们的工作将提供：一大组特征化、正交的 LSI-RDF 对；未来研究人员识别和/或设计 LSI-RDF 系统的新方法；推动进一步工程和新的光控制功能所需的结构信息和详细理解。PI 正在开发方法，向高中生介绍观看和玩弄 DNA 3D 结构的兴奋感，与社区团体合作讨论现代遗传工具的潜在力量，并鼓励大学 STEM 相关科目（尤其是女性和代表性不足的群体）入学，并与生物技术行业合作以改进课程 从事行业的学生。