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1.
ACS Synth Biol ; 3(12): 932-4, 2014 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-25408994

RESUMO

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.


Assuntos
Bioengenharia/métodos , Escherichia coli/metabolismo , Esterases/genética , Vetores Genéticos/genética , Mycobacterium smegmatis/metabolismo , Células Cultivadas , Escherichia coli/genética , Esterases/metabolismo , Vetores Genéticos/metabolismo , Humanos , Macrófagos/microbiologia
2.
ACS Synth Biol ; 3(12): 1003-6, 2014 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-25524110

RESUMO

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.


Assuntos
Bacteriófagos/genética , Resistência Microbiana a Medicamentos/genética , Escherichia coli/genética , Inativação Gênica , RNA Viral , Resistência Microbiana a Medicamentos/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Escherichia coli/metabolismo , Engenharia Genética , RNA Viral/genética , RNA Viral/farmacologia
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