In JoVE (1)
Articles by Aude G. Bernheim in JoVE
Phage-mediated Delivery of Targeted sRNA Constructs to Knock Down Gene Expression in E. coli Aude G. Bernheim*1, Vincent K. Libis*1, Ariel B. Lindner1, Edwin H. Wintermute1 1U1001, Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes We describe a method to knock down gene expression in a growing population of E. coli cells using sequence-targeted sRNA expression cassettes delivered by an M13 phagemid vector.
Other articles by Aude G. Bernheim on PubMed
In Situ Characterization of Mycobacterial Growth Inhibition by Lytic Enzymes Expressed in Vectorized E. Coli ACS Synthetic Biology. Dec, 2014 | Pubmed ID: 25408994 The emergence of extremely drug resistant Mycobacterium tuberculosis necessitates new strategies to combat the pathogen. Engineered bacteria may serve as vectors to deliver proteins to human cells, including mycobacteria-infected macrophages. In this work, we target Mycobacterium smegmatis, a nonpathogenic tuberculosis model, with E. coli modified to express trehalose dimycolate hydrolase (TDMH), a membrane-lysing serine esterase. We show that TDMH-expressing E. coli are capable of lysing mycobacteria in vitro and at low pH. Vectorized E. coli producing TDMH were found suppress the proliferation of mycobacteria in infected macrophages.
Silencing of Antibiotic Resistance in E. Coli with Engineered Phage Bearing Small Regulatory RNAs ACS Synthetic Biology. Dec, 2014 | Pubmed ID: 25524110 In response to emergent antibiotic resistance, new strategies are needed to enhance the effectiveness of existing antibiotics. Here, we describe a phagemid-delivered, RNA-mediated system capable of directly knocking down antibiotic resistance phenotypes. Small regulatory RNAs (sRNAs) were designed to specifically inhibit translation of chloramphenicol acetyltransferase and kanamycin phosphotransferase. Nonlytic phagemids coding for sRNA expression were able to infect and restore chloramphenicol and kanamycin sensitivity to populations of otherwise resistant E. coli. This modular system could easily be extended to other bacteria with resistance profiles that depend on specific transcripts.