Inhibition of gene expression with double strand RNA interference in Entamoeba histolytica

• Published in: Biochemical and biophysical research communications Vol. 320; no. 4; pp. 1118 - 1122
• Main Authors: Kaur, Gursharan, Lohia, Anuradha
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 06.08.2004
• Subjects: Animals; Cloning, Molecular - methods; Down-Regulation - genetics; dsRNA interference; Entamoeba histolytica; Entamoeba histolytica - genetics; Gene expression; Gene Expression Regulation - genetics; RNA Interference; RNA, Double-Stranded - genetics; Stable transformants; Transformation, Genetic - genetics; Entamoeba histolytica; Gene expression; dsRNA interference; Stable transformants
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2004.06.064

In order to inhibit gene expression in Entamoeba histolytica, we have developed a method based on expressing double strand RNA interference constructs in stable transformants. The 5′ end of Eh Dia was cloned head to head with an intervening non-specific stuffer fragment in the E. histolytica expression vector pJST4. This construct was transformed in E. histolytica HM1:IMSS trophozoites and stable transformants were selected with 20μg/ml G418. Our results show that expression of Eh Dia was completely inhibited in these transformants. These stable transformants could be maintained indefinitely without expression of Eh Dia. This method therefore provides an effective tool to study the phenotypic changes, which occur due to inhibition of gene expression in the absence of mutants and other microbiological manipulations in this protozoan parasite.