Antisense oligomer (ASO)-based antibiotics that target mRNAs of essential bacterial genes have great potential for counteracting antimicrobial resistance and for precision microbiome editing. To date, the development of such antisense antibiotics has primarily focused on using phosphorodiamidate morpholino (PMO) and peptide nucleic acid (PNA) backbones, largely ignoring the growing number of chemical modalities that have spurred the success of ASO-based human therapy. Here, we directly compare the activities of seven chemically distinct 10mer ASOs, all designed to target the essential gene acpP upon delivery with a KFF-peptide carrier into Salmonella. Our systematic analysis of PNA, PMO, phosphorothioate-modified DNA (PTO), 2’-methylated RNA (RNA-OMe), 2’-methoxyethylated RNA (RNA-MOE), 2’-fluorinated RNA (RNA-F) and 2’-4’-locked RNA (LNA) is based on a variety of in vitro and in vivo methods to evaluate ASO uptake, target pairing and inhibition of bacterial growth. Our data show that only PNA and PMO are efficiently delivered by the KFF peptide into Salmonella to inhibit bacterial growth. Nevertheless, the strong target binding affinity and in vitro translational repression activity of LNA and RNA-MOE make them promising modalities for antisense antibiotics that will require the identification of an effective carrier.

中文翻译：

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使用不同核苷酸模拟物的肽递送反义抗生素的比较分析

基于反义寡聚物 (ASO) 的抗生素以必需细菌基因的 mRNA 为目标，在对抗抗菌素耐药性和精准微生物组编辑方面具有巨大潜力。迄今为止，此类反义抗生素的开发主要集中在使用磷酸二酰胺吗啉 (PMO) 和肽核酸 (PNA) 主链，很大程度上忽略了越来越多的化学方式，这些方式促进了基于 ASO 的人类治疗的成功。在这里，我们直接比较了七种化学上不同的 10mer ASO 的活性，所有这些 ASO 都旨在通过 KFF 肽载体递送至沙门氏菌中后靶向必需基因 acpP。我们对 PNA、PMO、硫代磷酸修饰的 DNA (PTO)、2'-甲基化 RNA (RNA-OMe)、2'-甲氧基乙基化 RNA (RNA-MOE)、2'-氟化 RNA (RNA-F) 和 2 进行系统分析“-4”锁 RNA (LNA) 基于多种体外和体内方法来评估 ASO 摄取、靶标配对和细菌生长抑制。我们的数据表明，只有 PNA 和 PMO 能够通过 KFF 肽有效递送到沙门氏菌中以抑制细菌生长。然而，LNA 和 RNA-MOE 强大的靶标结合亲和力和体外翻译抑制活性使它们成为反义抗生素的有前途的模式，这需要鉴定有效的载体。