Adaptation of ribozyme technology to Leishmania

Summary

Principal Investigator: Nancy R Sturm
Abstract: DESCRIPTION (provided by the applicant): Directed ribozyme activity will add a valuable tool to the arsenal of techniques for study of small RNAs, and has not yet been reported in the kinetoplastids. This research is aimed at understanding gene expression in the family Trypanosomatidae, which includes the parasitic protozoan responsible for leishmaniasis, African sleeping sickness, and Chagas Disease. The focus is on the genesis and function of the spliced leader RNA, a small RNA which contributes the 5'-end sequence to every nuclear messenger RNA via a trans-splicing reaction. Trans-splicing is not found in the human host or insect vector, and thus represents a possible therapeutic target. This proposal outlines two series of experiments aimed at exploring the use of hammerhead ribozymes while elucidating the maturation pathway of the spliced leader RNA in Leishmania tarentolae: 1) Ribozyme activity in cis (mono-molecular) will be used to specify the 3' end of the spliced leader RNA. Stable products will allow us to overcome known processing defects for specific mutations in the spliced leader RNA and to examine downstream processing events including 5' methylation and trans-splicing. 2) Ribozymes will be used in trans (hi-molecular) to assay for nucleolar trafficking by attaching the ribozyme to a nucleolar U3 snoRNA. The nucleolar ribozyme will be directed against the spliced leader RNA and the spliced leader associated 1 RNA. Efficient ablation of RNA targets has great potential for elucidating the role of conserved sequences, structures and subcellular Iocalization. Once the parameters of the technique are established increasingly ambitious experiments can be designed including targeting of the spliced SL or discrete mRNAs.
Funding Period: 2003-04-01 - 2005-03-31
more information: NIH RePORT

Top Publications

  1. ncbi Human high density lipoproteins are platforms for the assembly of multi-component innate immune complexes
    April M Shiflett
    Josephine Bay Paul Center, Global Infectious Disease Program, Marine Biological Laboratory, Woods Hole, Massachussetts 02543, USA
    J Biol Chem 280:32578-85. 2005

Detail Information

Publications1

  1. ncbi Human high density lipoproteins are platforms for the assembly of multi-component innate immune complexes
    April M Shiflett
    Josephine Bay Paul Center, Global Infectious Disease Program, Marine Biological Laboratory, Woods Hole, Massachussetts 02543, USA
    J Biol Chem 280:32578-85. 2005
    ..These studies show that HDLs can serve as platforms for the assembly of multiple synergistic proteins and that these assemblies may play a critical role in the evolution of primate-specific innate immunity to trypanosome infection...