Literature DB >> 35085418

Growth and Genetic Manipulation of Entamoeba histolytica.

Rene L Suleiman1, Katherine S Ralston1.   

Abstract

Entamoeba histolytica is a parasitic protozoan and the causative agent of amoebiasis in humans. Amoebiasis has a high incidence of disease, resulting in ∼67,900 deaths per year, and it poses a tremendous burden of morbidity and mortality in children. Despite its importance, E. histolytica is an understudied parasite. These protocols describe the in vitro growth, maintenance, cryopreservation, genetic manipulation, and cloning of axenic E. histolytica trophozoites. There has been significant progress in genetic manipulation of this organism over the past decade, and these protocols outline the ways in which these advances can be implemented.
© 2022 Wiley Periodicals LLC. Basic Protocol 1: Culturing E. histolytica trophozoites Support Protocol 1: Preparation of TYI-S-33 medium Support Protocol 2: Lot testing of Biosate peptone and adult bovine serum for TYI-S-33 medium Basic Protocol 2: Cryopreservation of E. histolytica trophozoites Support Protocol 3: Preparation of cryoprotectant solutions Basic Protocol 3: Transfection of E. histolytica trophozoites with Attractene reagent Basic Protocol 4: Creating clonal lines using limiting dilution Basic Protocol 5: Knockdown of one to two genes with trigger-induced RNA interference Support Protocol 4: Evaluation of RNA interference knockdown with reverse transcriptase PCR Basic Protocol 6: E. histolytica growth curves. © 2022 Wiley Periodicals LLC.

Entities:  

Keywords:  RNAi; cryopreservation; culture; entamoeba histolytica; parasite; transfection

Mesh:

Substances:

Year:  2022        PMID: 35085418      PMCID: PMC9040564          DOI: 10.1002/cpz1.327

Source DB:  PubMed          Journal:  Curr Protoc        ISSN: 2691-1299


  40 in total

1.  Nucleus-localized antisense small RNAs with 5'-polyphosphate termini regulate long term transcriptional gene silencing in Entamoeba histolytica G3 strain.

Authors:  Hanbang Zhang; Hussein Alramini; Vy Tran; Upinder Singh
Journal:  J Biol Chem       Date:  2011-11-02       Impact factor: 5.157

2.  Involvement of a short interspersed element in epigenetic transcriptional silencing of the amoebapore gene in Entamoeba histolytica.

Authors:  Michael Anbar; Rivka Bracha; Yael Nuchamowitz; Yan Li; Anat Florentin; David Mirelman
Journal:  Eukaryot Cell       Date:  2005-11

3.  Stable transfection of Entamoeba histolytica trophozoites by lipofection.

Authors:  A Olvera; F Olvera; R R Vines; F Recillas-Targa; P M Lizardi; S Dhar; S Bhattacharya; W Petri; A Alagón
Journal:  Arch Med Res       Date:  1997       Impact factor: 2.235

4.  A tetracycline-inducible gene expression system in Entamoeba histolytica.

Authors:  G Ramakrishnan; R R Vines; B J Mann; W A Petri
Journal:  Mol Biochem Parasitol       Date:  1997-01       Impact factor: 1.759

5.  Transient transfection of the enteric parasite Entamoeba histolytica and expression of firefly luciferase.

Authors:  J E Purdy; B J Mann; L T Pho; W A Petri
Journal:  Proc Natl Acad Sci U S A       Date:  1994-07-19       Impact factor: 11.205

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

Authors:  Gursharan Kaur; Anuradha Lohia
Journal:  Biochem Biophys Res Commun       Date:  2004-08-06       Impact factor: 3.575

7.  Improved method for the monoxenic cultivation of Entamoeba histolytica Schaudinn, 1903 and E. histolytica-like amebae with trypanosomatids.

Authors:  L S Diamond
Journal:  J Parasitol       Date:  1968-08       Impact factor: 1.276

8.  Agarose gel electrophoresis.

Authors:  D Voytas
Journal:  Curr Protoc Mol Biol       Date:  2001-05

9.  Transfection and continuous expression of heterologous genes in the protozoan parasite Entamoeba histolytica.

Authors:  L Hamann; R Nickel; E Tannich
Journal:  Proc Natl Acad Sci U S A       Date:  1995-09-12       Impact factor: 11.205

10.  Transcriptional silencing of multiple genes in trophozoites of Entamoeba histolytica.

Authors:  Rivka Bracha; Yael Nuchamowitz; Michael Anbar; David Mirelman
Journal:  PLoS Pathog       Date:  2006-05-26       Impact factor: 6.823

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  1 in total

1.  Entamoeba histolytica Develops Resistance to Complement Deposition and Lysis after Acquisition of Human Complement-Regulatory Proteins through Trogocytosis.

Authors:  Hannah W Miller; Tammie S Y Tam; Katherine S Ralston
Journal:  mBio       Date:  2022-03-01       Impact factor: 7.786

  1 in total

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