Banca de DEFESA: Alessandro Zanard Lopes Ferreira

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : Alessandro Zanard Lopes Ferreira
DATE: 17/04/2023
TIME: 09:00
LOCAL: Plataforma Teams
TITLE:

Functional Analysis of Kinesin C from Trypanosoma Cruzi using the CRISPR/Cas9 system

KEY WORDS:

Chagas Disease; Trypanosoma cruzi; CRISPR/Cas9; Kinesin.

PAGES: 116
BIG AREA: Ciências Biológicas
AREA: Genética
SUMMARY:

Chagas disease (CD) is a parasitic disease caused by the protozoan Trypanosoma cruzi. Considered a neglected tropical disease, CD affects millions of people in the 21 Latin American countries where it is endemic and in the countries receiving immigrants from that region. Due to the scarcity of drugs available for its treatment, there is great interest in knowing the homeostatic components of the trypanosomatid Trypanosoma cruzi, in order to seek possible pharmacological targets. In this sense, considering the already known importance of kinesins in several diseases due to their role in the regulation of cytokinesis and maintenance of cell morphology, we sought in this work to characterize the motor protein kinesin C (TcKIN-C) of T. cruzi and its importance in homeostasis cell phone. Initially, an in silico survey of the characteristics of TcKIN-C, an orphan kinesin present only in T. cruzi, expressed by the TcCLB.504427.260 and TcCLB.508043.40 alleles, was carried out. Additionally, using the premise of metagenomics, the CRISPR/Cas9 biotechnological tool was used in order to cause an interruption in TcKIN-C expression, resulting in truncated expression of this protein and possible loss of its functionality. Through the in silico analysis, a significant difference was observed in the sequence of the two alleles, so an analysis of both alleles was performed using bioinformatics tools, from alignments, prediction of post-translational modifications (MPTs), interaction networks to the 3D structure of both alleles. Then, a knockout was performed using the aforementioned technique, where two strategies were outlined: an initial knockout (called Strategy I) without selection to observe the effects at the population level and a second knockout (called Strategy II), where the transgenic population was selected and the effects of TcKIN-C deletion were studied in more detail. As of the date of this publication, no other work has been published that makes the functional characterization of a kinesin in Trypanosoma cruzi. In short, as observed in Trypanosoma brucei, this work corroborates the notion that kinesins are of central importance for the survival of these parasites. Furthermore, it was also possible to achieve the objective of selecting transgenic populations, allowing further studies.

BANKING MEMBERS:
Externa à Instituição - Clênia dos Santos Azevedo - MS
Presidente - 2492352 - IZABELA MARQUES DOURADO BASTOS CHARNEAU
Externa à Instituição - PAULA BEATRIZ DE MEDEIROS SANTIAGO - UnB
Externo ao Programa - 1876284 - VICENTE DE PAULO MARTINS