RNAi libraries and kinetoplast DNA

Englund, Paul T.; Agbo, E.E.C.; Lindsay, Megan E.; Liu, B.; Liu, Y.; Motyka, Shawn A.; Yildirir, Gokben; Zhao, Zhixing

doi:10.1042/bst20051409

Cited by 6 publications

(2 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, the research group of Paul Englund described a novel method for conducting forward genetic screens in T. brucei [27, 89]. This method employs a tetracycline-inducible RNAi library that contains random T. brucei DNA fragments between two opposing, tetracycline-inducible promoters in a plasmid that can be stably integrated into the genome (Fig.…”

Section: Methodsmentioning

confidence: 99%

Approaches for Functional Analysis of Flagellar Proteins in African Trypanosomes

Oberholzer

López

Ralston

et al. 2009

Methods in Cell Biology

View full text Add to dashboard Cite

The eukaryotic flagellum is a highly conserved organelle serving motility, sensory and transport functions. Although genetic, genomic and proteomic studies have led to the identification of hundreds of flagellar and putative flagellar proteins, precisely how these proteins function individually and collectively to drive flagellum motility and other functions remains to be determined. In this chapter we provide an overview of tools and approaches available for studying flagellum protein function in the protozoan parasite Trypanosoma brucei. We begin by outlining techniques for in vitro cultivation of both T. brucei lifecycle stages, as well as transfection protocols for the delivery of DNA constructs. We then describe specific assays used to assess flagellum function including flagellum preparation and quantitative motility assays. We conclude the chapter with a description of molecular genetic approaches for manipulating gene function. In summary, the availability of potent molecular tools, as well as the health and economic relevance of T. brucei as a pathogen, combine to make the parasite an attractive and integral experimental system for the functional analysis of flagellar proteins.

show abstract

Section: Methodsmentioning

confidence: 99%

Approaches for Functional Analysis of Flagellar Proteins in African Trypanosomes

Oberholzer

López

Ralston

et al. 2009

Methods in Cell Biology

View full text Add to dashboard Cite

show abstract

“…For example, efficient homologous recombination facilitates GFP-fusions for the examination of the cellular location of proteins [65], while inducible RNAi enables knockdown analysis to examine their function [75,76]. Genome-wide RNAi libraries are available [77,78,98]. Furthermore, the organism's doubling time is 6–9 h (cf.…”

Section: Trypanosoma Brucei As a Model To Study Chromosome Segregationmentioning

confidence: 99%

Evolutionary cell biology of chromosome segregation: insights from trypanosomes

Akiyoshi

Gull

2013

Open Biol.

View full text Add to dashboard Cite

Faithful transmission of genetic material is essential for the survival of all organisms. Eukaryotic chromosome segregation is driven by the kinetochore that assembles onto centromeric DNA to capture spindle microtubules and govern the movement of chromosomes. Its molecular mechanism has been actively studied in conventional model eukaryotes, such as yeasts, worms, flies and human. However, these organisms are closely related in the evolutionary time scale and it therefore remains unclear whether all eukaryotes use a similar mechanism. The evolutionary origins of the segregation apparatus also remain enigmatic. To gain insights into these questions, it is critical to perform comparative studies. Here, we review our current understanding of the mitotic mechanism in Trypanosoma brucei, an experimentally tractable kinetoplastid parasite that branched early in eukaryotic history. No canonical kinetochore component has been identified, and the design principle of kinetochores might be fundamentally different in kinetoplastids. Furthermore, these organisms do not appear to possess a functional spindle checkpoint that monitors kinetochore–microtubule attachments. With these unique features and the long evolutionary distance from other eukaryotes, understanding the mechanism of chromosome segregation in T. brucei should reveal fundamental requirements for the eukaryotic segregation machinery, and may also provide hints about the origin and evolution of the segregation apparatus.

show abstract

Cell cycle regulation in Trypanosoma brucei

Hammarton

2007

Molecular and Biochemical Parasitology

133

144

View full text Add to dashboard Cite

Cell division is regulated by intricate and interconnected signal transduction pathways that precisely coordinate, in time and space, the complex series of events involved in replicating and segregating the component parts of the cell. In Trypanosoma brucei, considerable progress has been made over recent years in identifying molecular regulators of the cell cycle and elucidating their functions, although many regulators undoubtedly remain to be identified, and there is still a long way to go with respect to determining signal transduction pathways. However, it is clear that cell cycle regulation in T. brucei is unusual in many respects. Analyses of trypanosome orthologues of conserved eukaryotic cell cycle regulators have demonstrated divergence of their function in the parasite, and a number of other key regulators are missing from T. brucei. Cell cycle regulation differs in different parasite life cycle stages, and T. brucei appears to use different checkpoint control strategies compared to model eukaryotes. It is therefore probable that T. brucei has evolved novel pathways to control its cell cycle.

show abstract

RNAi libraries and kinetoplast DNA

Cited by 6 publications

References 23 publications

Approaches for Functional Analysis of Flagellar Proteins in African Trypanosomes

Approaches for Functional Analysis of Flagellar Proteins in African Trypanosomes

Evolutionary cell biology of chromosome segregation: insights from trypanosomes

Cell cycle regulation in Trypanosoma brucei

Contact Info

Product

Resources

About