Inhibition of gene expression with double strand RNA interference in Entamoeba histolytica
- PMID: 15249205
- DOI: 10.1016/j.bbrc.2004.06.064
Inhibition of gene expression with double strand RNA interference in Entamoeba histolytica
Abstract
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 20microg/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.
Similar articles
-
Double-stranded RNA mediates homology-dependent gene silencing of gamma-tubulin in the human parasite Entamoeba histolytica.Mol Biochem Parasitol. 2004 Nov;138(1):21-8. doi: 10.1016/j.molbiopara.2004.07.005. Mol Biochem Parasitol. 2004. PMID: 15500912
-
Silencing genes by RNA interference in the protozoan parasite Entamoeba histolytica.Methods Mol Biol. 2008;442:113-28. doi: 10.1007/978-1-59745-191-8_9. Methods Mol Biol. 2008. PMID: 18369782
-
Molecular characterization of Entamoeba histolytica RNase III and AGO2, two RNA interference hallmark proteins.Exp Parasitol. 2005 Jul;110(3):265-9. doi: 10.1016/j.exppara.2005.02.023. Epub 2005 Apr 7. Exp Parasitol. 2005. PMID: 15955322
-
Gene therapy progress and prospects. Downregulating gene expression: the impact of RNA interference.Gene Ther. 2004 Aug;11(16):1241-8. doi: 10.1038/sj.gt.3302324. Gene Ther. 2004. PMID: 15292914 Review.
-
RNA interference in protozoan parasites.Cell Microbiol. 2004 Jun;6(6):509-19. doi: 10.1111/j.1462-5822.2004.00399.x. Cell Microbiol. 2004. PMID: 15104593 Review.
Cited by
-
Long double-stranded RNA produces specific gene downregulation in Giardia lamblia.J Parasitol. 2010 Aug;96(4):815-9. doi: 10.1645/GE-2406.1. J Parasitol. 2010. PMID: 20476805 Free PMC article.
-
Entamoeba histolytica: a snapshot of current research and methods for genetic analysis.Curr Opin Microbiol. 2012 Aug;15(4):469-75. doi: 10.1016/j.mib.2012.04.011. Epub 2012 Jun 2. Curr Opin Microbiol. 2012. PMID: 22664276 Free PMC article. Review.
-
Silencing of Entamoeba histolytica glucosamine 6-phosphate isomerase by RNA interference inhibits the formation of cyst-like structures.Biomed Res Int. 2013;2013:758341. doi: 10.1155/2013/758341. Epub 2013 Jan 22. Biomed Res Int. 2013. PMID: 23484154 Free PMC article.
-
Short hairpin RNA-mediated knockdown of protein expression in Entamoeba histolytica.BMC Microbiol. 2009 Feb 17;9:38. doi: 10.1186/1471-2180-9-38. BMC Microbiol. 2009. PMID: 19222852 Free PMC article.
-
RNA interference in Entamoeba histolytica: implications for parasite biology and gene silencing.Future Microbiol. 2011 Jan;6(1):103-17. doi: 10.2217/fmb.10.154. Future Microbiol. 2011. PMID: 21162639 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
