融合脂质体将寡核苷酸递送至细菌中。

Delivery of Oligonucleotides into Bacteria by Fusogenic Liposomes.

机构信息

LEPABE - Laboratory for Process Engineering, Environment, Biotechnology and Energy, Department of Chemical Engineering, Faculty of Engineering, University of Porto, Porto, Portugal.

Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Ghent, Belgium.

出版信息

Methods Mol Biol. 2021;2246:87-96. doi: 10.1007/978-1-0716-1115-9_6.

DOI:10.1007/978-1-0716-1115-9_6

PMID:33576984

Abstract

Oligonucleotides able to hybridize bacterial RNA via in situ hybridization may potentially act as new antimicrobials, replacing antibiotics, and as fast in vivo diagnostic probes, outperforming current clinical methodologies. Nonetheless, oligonucleotides are not able to efficiently permeate the multi-layered bacterial envelope to reach their target RNA in the cytosol. Cationic fusogenic liposomes are here suggested as vehicles to enable the internalization of oligonucleotides in bacteria. Here, we describe the formulation of DOTAP-DOPE liposomes, their complexation with small negatively charged oligonucleotides, and the evaluation of the intracellular delivery of the oligonucleotides in bacteria. This strategy uncovers the potential of performing FISH in vivo for real-time detection and treatment of infections.

摘要

通过原位杂交与细菌 RNA 杂交的寡核苷酸可能潜在地作为新型抗菌药物替代抗生素，同时也作为快速的体内诊断探针，优于当前的临床方法。然而，寡核苷酸无法有效地穿透多层细菌包膜，到达细胞质中的靶 RNA。阳离子融合脂质体被提议作为载体，使寡核苷酸在细菌中内化。在此，我们描述了 DOTAP-DOPE 脂质体的配方、它们与带负电荷的小寡核苷酸的复合物的形成，以及在细菌中寡核苷酸的细胞内递释的评估。该策略揭示了在体内进行 FISH 的潜力，以实时检测和治疗感染。