Inhibition of gene expression in Entamoeba histolytica with antisense peptide nucleic acid oligomers.
Peptide nucleic acids (PNAs) may be a potent tool for gene function studies in medically important parasitic organisms, especially those that have not before been accessible to molecular genetic knockout approaches. One such organism is Entamoeba histolytica, the causative agent of amebiasis, which infects about 500 million people and is the cause of clinical disease in over 40 million each year, mainly in the tropical and subtropical world. We used PNA antisense oligomers to inhibit expression of an episomally expressed gene (neomycin phosphorotransferase, NPT) and a chromosomal gene (EhErd2, a homolog of Erd2, a marker of the Golgi system in eukaryotic cells) in axenically cultured trophozoites of E. histolytica. Measurement of NPT enzyme activity and EhErd2 protein levels, as well as measurement of cellular proliferation, revealed specific decreases in expression of the target genes, and concomitant inhibition of cell growth, in trophozoites treated with micromolar concentrations of unmodified antisense PNA oligomers.
Instituto de Biotecnología/UNAM. Av. Universidad 2001, 62210 Morelos, Mexico.
R P
RP
Stock
A
A
Olvera
R
R
Sánchez
A
A
Saralegui
S
S
Scarfì
R
R
Sanchez-Lopez
M A
MA
Ramos
L C
LC
Boffa
U
U
Benatti
A
A
Alagón
D000818:Animals; D016375:Antisense Elements (Genetics) / Q000235:genetics / Q000494:pharmacology*; D015415:Biomarkers / Q000032:analysis; D002455:Cell Division / Q000187:drug effects; D015536:Down-Regulation / Q000187:drug effects*; D004748:Entamoeba histolytica / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D005839:Gentamicins / Q000494:pharmacology; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D019868:Kanamycin Kinase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009355:Neomycin / Q000378:metabolism; D020135:Peptide Nucleic Acids / Q000235:genetics / Q000494:pharmacology*; D010539:Permeability; D014162:Transfection
D016375:Antisense Elements (Genetics); D015415:Biomarkers; D005839:Gentamicins; D008565:Membrane Proteins; D020135:Peptide Nucleic Acids; C010680:antibiotic G 418; D019868:Kanamycin Kinase; D009355:Neomycin