Monitoring processing, localisation and function of differentially expressed ncRNAs in Leishmania braziliensis by multi-colour RNA FISH on subcellular resolution

• Grant number: 21/15182-3
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: April 01, 2022
• End date: March 31, 2023
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Angela Kaysel Cruz
• Grantee: José Carlos Quilles Junior
• Supervisor: Susanne Kramer
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: Universität Würzburg, Germany
• Associated to the scholarship: 20/00088-9 - Epigenetic regulation of Leishmania gene expression, BP.PD

Abstract

The discovery of non-coding RNAs (ncRNAs) that act as regulatory elements has significantly contributed to the development and control of many aspects from cellular and molecular points of view. In Leishmania protozoa, ncRNAs are still poorly explored, particularly at the functional level. The genetic organization of these parasites transfer to the post-transcriptional level the control of gene expression which is not only, but heavily based on mRNA fate and stability, translation activity and protein degradation modulation. We have previously identified a large number of ncRNAs differentially expressed (DE ncRNAs) during the life cycle of L. braziliensis, suggesting that such ncRNAs may play relevant functions in a specific parasite life stage. To investigate putative regulatory function, we selected 50 DE ncRNAs among those predicted in silico, based on the level of differential expression. They include intergenic transcripts and located in Switch Strand Region (SSR). For ten of them, we confirmed they exist as independent transcripts and phenotypes linked to the ncRNA knockout or overexpression are in course (as presented in the accompanying scientific report from Brazil). Taking all these relevant ncRNA information in L. braziliensis, we intend here to investigate the traffic and stability of ten RNA species inside the parasite cell by dual colour RNA FISH using confocal microscopy, exploring their behavior and intracellular location in different parasite morphologies and under specific environmental conditions, in order to better understand their real function. At the end, we hope to join relevant information about this class of RNA in order to elucidate how stable they are and to where they go inside the cell, a study not reported in Leishmania until now. (AU)