Naegleria fowleri: nfa1 gene knock-down by double-stranded RNAs

• Published in: Experimental parasitology 2008-02, Vol.118 (2), p.208-213
• Main Authors: Jung, Suk-Yul, Kim, Jong-Hyun, Lee, Yang-Jin, Song, Kyoung-Ju, Kim, Kyongmin, Park, Sun, Im, Kyung-il, Shin, Ho-Joon
• Format: Article
• Language: English
• Subjects: Animals; Antigens, Protozoan - genetics; Antigens, Protozoan - physiology; Biological and medical sciences; Blotting, Northern; Blotting, Southern; Blotting, Western; Double-stranded RNA; Electrophoresis, Agar Gel; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation; Gene knock-down; Life cycle. Host-agent relationship. Pathogenesis; Naegleria fowleri; Naegleria fowleri - genetics; nfa1 gene; Protozoa; Protozoan Proteins - genetics; Protozoan Proteins - physiology; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference - physiology; RNA, Double-Stranded - genetics; Transfection
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2007.08.008

Nfa1 protein expressed by the nfa1 gene that was cloned recently from pathogenic Naegleria fowleri was found in pseudopodia, especially food-cups, and concerned with a mechanism of pathogenicity of N. fowleri. In the present study, N. fowleri nfa1 gene was knocked down using double-stranded RNAs, and the expression of Nfa1 protein was observed. Using synthetic double-stranded RNA of the nfa1 gene in vitro, the nfa1 gene and Nfa1 protein were knocked down about 50.4 ± 3.1% and 52 ± 2%, respectively. These results suggest that RNA interference (RNAi) may be an effective technique for gene knock-down in N. fowleri trophozoites.