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High-throughput Gene Tagging in Trypanosoma brucei
Published on: August 12, 2016
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Genome-scale RNA interference profiling of Trypanosoma brucei cell cycle progression defects
Catarina A Marques1,2, Melanie Ridgway1, Michele Tinti1
1Wellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, UK.
Nature Communications
|September 10, 2022
Summary
This study screens for cell cycle defects in Trypanosoma brucei, revealing over 100 flagellar components affecting genome replication and identifying metabolic and regulatory controls crucial for cell division.
Area of Science:
- Molecular Biology
- Parasitology
- Genomics
Background:
- Trypanosomatids are significant human and livestock pathogens.
- Cell cycle control mechanisms in these protozoa remain poorly understood.
Purpose of the Study:
- To perform a genome-wide RNA-interference screen for cell cycle defects in Trypanosoma brucei.
- To identify genes and pathways regulating trypanosome cell cycle progression.
Main Methods:
- Genome-wide RNA-interference library screen in Trypanosoma brucei.
- High-throughput flow cytometry for cell sorting.
- Deep sequencing of RNAi targets.
- Digital reconstruction of cell cycle profiles.
- Online data visualization tool (tryp-cycle.pages.dev).
Main Results:
- >100 flagellar component knockdowns linked to genome endoreduplication.
- Evidence for metabolic regulation of the G1-S transition.
- Surface antigen regulatory mRNA-binding protein knockdowns associated with G2M accumulation.
- Identification of a nucleoredoxin involved in mitochondrial genome segregation and mitosis.
Conclusions:
- Provides comprehensive functional genomic data on trypanosome cell cycle regulation.
- Identifies novel and known regulators coordinating cell division in Trypanosoma brucei.
- Highlights the link between flagellar function and genome stability.
