Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System

10.3791/59962 ◽  
2019 ◽  
Author(s):  
Yukie Akutsu ◽  
Motomichi Doi ◽  
Koji Furukawa ◽  
Yuko Takagi
Keyword(s):  
Trypanosoma Cruzi ◽  
Gene Knockout ◽  
Amastigote Stage

scholarly journals Amaryllidaceae alkaloids with anti-Trypanosoma cruzi activity

2020 ◽  
Author(s):  
Nieves Martinez-Peinado ◽  
Nuria Cortes-Serra ◽  
Laura Torras-Claveria ◽  
Maria-Jesus Pinazo ◽  
Joaquim Gascon ◽  
...  
Keyword(s):  
Natural Products ◽  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Specific Activity ◽  
New Drugs ◽  
Host Cells ◽  
Amastigote Stage ◽  
Limited Efficacy ◽  
Single Compound ◽  
New Compounds

Abstract Background: Chagas disease, caused by the protozoan Trypanosoma cruzi, is a neglected disease that affects ~7 million people worldwide. Development of new drugs to treat the infection remains a priority since those currently available have frequent side effects and limited efficacy at the chronic stage. Natural products provide a pool of diversity structures to lead the chemical synthesis of novel molecules for this purpose. Herein we analyzed the anti- T. cruzi activity of 9 alkaloids derived from plants of the Amaryllidaceae family. Methods: the activity of each alkaloid was assessed by means of an anti- T. cruzi phenotypic assay. We further evaluated the compounds that inhibited the parasite growth on two distinct cytotoxicity assays to discard those that were toxic to host cells and assure parasite selectivity. Results: we identified a single compound (hippeastrine 2 ) that was selectively active against the parasite yielding selectivity indexes of 12.7 and 35.2 against Vero and HepG2 cells, respectively. Moreover, it showed specific activity against the amastigote stage (IC 50 = 3.31 μM). Conclusions: results reported here suggest that natural products are an interesting source of new compounds for the development of drugs against Chagas disease.

scholarly journals Evaluation of high efficiency gene knockout strategies for Trypanosoma cruzi

2009 ◽  
Vol 9 (1) ◽  
pp. 90 ◽  
Author(s):  
Dan Xu ◽  
Cecilia Pérez Brandán ◽  
Miguel Basombrío ◽  
Rick L Tarleton
Keyword(s):  
Trypanosoma Cruzi ◽  
High Efficiency ◽  
Gene Knockout

scholarly journals gp63 Homologues in Trypanosoma cruzi: Surface Antigens with Metalloprotease Activity and a Possible Role in Host Cell Infection

2003 ◽  
Vol 71 (10) ◽  
pp. 5739-5749 ◽  
Author(s):  
Ileana C. Cuevas ◽  
Juan J. Cazzulo ◽  
Daniel O. Sánchez
Keyword(s):  
Trypanosoma Cruzi ◽  
Genomic Organization ◽  
Vero Cells ◽  
Surface Proteins ◽  
Cell Infection ◽  
Northern Blots ◽  
Western Blots ◽  
Amastigote Stage ◽  
Group Members ◽  
Metalloprotease Activity

ABSTRACT gp63 is a highly abundant glycosylphosphatidylinositol (GPI)-anchored membrane protein expressed predominantly in the promastigote but also in the amastigote stage of Leishmania species. In Leishmania spp., gp63 has been implicated in a number of steps in establishment of infection. Here we demonstrate that Trypanosoma cruzi, the etiological agent of Chagas' disease, has a family of gp63 genes composed of multiple groups. Two of these groups, Tcgp63-I and -II, are present as high-copy-number genes. The genomic organization and mRNA expression pattern were specific for each group. Tcgp63-I was widely expressed, while the Tcgp63-II group was scarcely detected in Northern blots, even though it is well represented in the T. cruzi genome. Western blots using sera directed against a synthetic peptide indicated that the Tcgp63-I group produced proteins of ∼78 kDa, differentially expressed during the life cycle. Immunofluorescence staining and phosphatidylinositol-specific phospholipase C digestion confirmed that Tcgp63-I group members are surface proteins bound to the membrane by a GPI anchor. We also demonstrate the presence of metalloprotease activity which is attributable, at least in part, to Tcgp63-I group. Since antibodies against Tcgp63-I partially blocked infection of Vero cells by trypomastigotes, a possible role for this group in infection is suggested.

scholarly journals Amaryllidaceae alkaloids with anti-Trypanosoma cruzi activity

2020 ◽  
Author(s):  
Nieves Martinez-Peinado ◽  
Nuria Cortes-Serra ◽  
Laura Torras-Claveria ◽  
Maria-Jesus Pinazo ◽  
Joaquim Gascon ◽  
...  
Keyword(s):  
Natural Products ◽  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Specific Activity ◽  
New Drugs ◽  
Host Cells ◽  
Amastigote Stage ◽  
Limited Efficacy ◽  
Single Compound ◽  
New Compounds

Abstract Background: Chagas disease, caused by the protozoan Trypanosoma cruzi, is a neglected disease that affects ~7 million people worldwide. Development of new drugs to treat the infection remains a priority since those currently available have frequent side effects and limited efficacy at the chronic stage. Natural products provide a pool of diversity structures to lead the chemical synthesis of novel molecules for this purpose. Herein we analyzed the anti- T. cruzi activity of 9 alkaloids derived from plants of the Amaryllidaceae family. Methods: the activity of each alkaloid was assessed by means of an anti- T. cruzi phenotypic assay. We further evaluated the compounds that inhibited the parasite growth on two distinct cytotoxicity assays to discard those that were toxic to host cells and assure parasite selectivity. Results: we identified a single compound (hippeastrine 2 ) that was selectively active against the parasite yielding selectivity indexes of 12.7 and 35.2 against Vero and HepG2 cells, respectively. Moreover, it showed specific activity against the amastigote stage (IC 50 = 3.31 μM). Conclusions: results reported here suggest that natural products are an interesting source of new compounds for the development of drugs against Chagas disease.

Use of Antigen Preparations of the Amastigote Stage of Trypanosoma cruzi in the Serology of Chagas' Disease *

1984 ◽  
Vol 33 (3) ◽  
pp. 362-371 ◽  
Author(s):  
Fausto G. Araujo ◽  
Douglas Guptill
Keyword(s):  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Amastigote Stage

scholarly journals IL-10-IFN-γ Double Producers CD4+ T Cells Are Induced by Immunization with an Amastigote Stage Specific Derived Recombinant Protein of Trypanosoma Cruzi

2011 ◽  
Vol 7 (8) ◽  
pp. 1093-1100 ◽  
Author(s):  
Yevel Flores-García ◽  
José Luis Rosales-Encina ◽  
Abhay R. Satoskar ◽  
Patricia Talamás-Rohana
Keyword(s):  
T Cells ◽  
Recombinant Protein ◽  
Trypanosoma Cruzi ◽  
Cd4 T Cells ◽  
Amastigote Stage ◽  
Ifn Γ

Assessment of two CRISPR-Cas9 genome editing protocols for rapid generation of Trypanosoma cruzi gene knockout mutants

2018 ◽  
Vol 48 (8) ◽  
pp. 591-596 ◽  
Author(s):  
Gabriela Assis Burle-Caldas ◽  
Melissa Soares-Simões ◽  
Laiane Lemos-Pechnicki ◽  
Wanderson Duarte DaRocha ◽  
Santuza M.R. Teixeira
Keyword(s):  
Trypanosoma Cruzi ◽  
Genome Editing ◽  
Gene Knockout ◽  
Knockout Mutants ◽  
Rapid Generation

scholarly journals Amaryllidaceae alkaloids with anti-Trypanosoma cruzi activity.

2020 ◽  
Author(s):  
Nieves Martinez-Peinado ◽  
Nuria Cortes-Serra ◽  
Laura Torras-Claveria ◽  
Maria-Jesus Pinazo ◽  
Joaquim Gascon ◽  
...  
Keyword(s):  
Natural Products ◽  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Specific Activity ◽  
New Drugs ◽  
Host Cells ◽  
Amastigote Stage ◽  
Limited Efficacy ◽  
Single Compound ◽  
New Compounds

Abstract Background: Chagas disease, caused by the protozoan Trypanosoma cruzi, is a neglected disease that affects ~7 million people worldwide. Development of new drugs to treat the infection remains a priority since those currently available have frequent side effects and limited efficacy at the chronic stage. Natural products provide a pool of diversity structures to lead the chemical synthesis of novel molecules for this purpose. Herein we analyzed the anti-T. cruzi activity of 9 alkaloids derived from plants of the Amaryllidaceae family.Methods: the activity of each alkaloid was assessed by means of an anti-T. cruzi phenotypic assay. We further evaluated the compounds that inhibited the parasite growth on two distinct cytotoxicity assays to discard those that were toxic to host cells and assure parasite selectivity.Results: we identified a single compound (hippeastrine 2) that was selectively active against the parasite yielding selectivity indexes of 12.7 and 35.2 against Vero and HepG2 cells, respectively. Moreover, it showed specific activity against the amastigote stage (IC50 = 3.31 μM).Conclusions: results reported here suggest that natural products are an interesting source of new compounds for the development of drugs against Chagas disease.

Protective effect of Trypanosoma rangeli against infections with a highly virulent strain of Trypanosoma cruzi

1997 ◽  
Vol 2 (5) ◽  
pp. 482-487 ◽  
Author(s):  
Claudio Zuniga ◽  
Teresa Palau ◽  
Pilar Penin ◽  
Carlos Gamallo ◽  
Jose Antonio de Diego
Keyword(s):  
Trypanosoma Cruzi ◽  
Protective Effect ◽  
Virulent Strain ◽  
Trypanosoma Rangeli ◽  
Highly Virulent
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