细菌转染,一种细菌介导的遗传转移形式,被强调为基因治疗的替代机制。用于免疫反应性目的的该系统的关键优势源于能够通过吸引抗原呈递细胞(APC)的识别和细胞摄取来启动免疫应答的细菌宿主的性质。该方法也是更有效地递送较大遗传构建体的合适技术,因为它可以将不同大小的质粒转移到靶哺乳动物细胞中。鉴于这些优势,正在研究细菌载体作为将质粒DNA递送到靶细胞中以实现异源蛋白表达的潜在载体。用于细菌转染的细菌通常是非致病性的;然而,由于使用生物制剂而引起的关注。为了克服这些担忧,增强的细菌降解已被设计为细菌转染载体的减毒和安全特征。特别是,ΦX174裂解E(LyE)基因可以被重新利用以最小化哺乳动物宿主内的细菌存活,同时还改善整体基因递送。更具体地说,当使用鼠RAW264.7巨噬细胞APC测试时,携带LyE基因的工程化细菌载体显示出改善的基因递送和安全性。本章概述了在疫苗应用背景下,将大肠杆菌作为一种更安全、更有效的遗传抗原递送杆菌转染载体进行LyE表达所采取的步骤。
Bactofection, a bacterial-mediated form of genetic transfer, is highlighted as an alternative mechanism for gene therapy. A key advantage of this system for immune-reactivity purposes stems from the nature of the bacterial host capable of initiating an immune response by attracting recognition and cellular uptake by antigen-presenting cells (APCs). The approach is also a suitable technique to deliver larger genetic constructs more efficiently as it can transfer plasmids of varying sizes into target mammalian cells. Given these advantages, bacterial vectors are being studied as potential carriers for the delivery of plasmid DNA into target cells to enable expression of heterologous proteins. The bacteria used for
bactofection are generally nonpathogenic; however, concerns arise due to the use of a biological agent. To overcome such concerns, enhanced bacterial degradation has been engineered as an attenuation and safety feature for
bactofection vectors. In particular, the ΦX174 lysis E (LyE) gene can be repurposed to both minimize bacterial survival within mammalian hosts while also improving overall gene delivery. More specifically, an engineered bacterial vector carrying the LyE gene showed improved gene delivery and safety profiles when tested with murine RAW264.7 macrophage APCs. This chapter outlines steps taken to engineer E. coli for LyE expression as a safer and more effective genetic antigen delivery
bactofection vehicle in the context of vaccine utility.