Development of an sRNA-mediated conditional knockdown system for <i>Chlamydia trachomatis</i>.

ABSTRACT: We describe a new Chlamydia trachomatis protein depletion method that uses an engineered small RNA (sRNA) to inhibit translation of a target gene. In proof-of-principle experiments, we induced functional knockdown of IncA, a fusion-mediating inclusion membrane protein, as shown with Western blots, loss of IncA staining at the inclusion membrane, and production of multiple chlamydial inclusions within an infected cell. These effects were titratable and reversible. To test for polar effects, we separately targeted the inclusion membrane proteins IncE and IncG, which are expressed from the incDEFG operon. Knockdown of IncE caused loss of IncE and its interacting host protein SNX6 at the inclusion membrane, without affecting IncG protein levels. Similarly, IncG knockdown signific

SUBMITTER: Ehses J 

PROVIDER: S-EPMC11796381 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature