Epigenetic regulation of Leishmania gene expression

• Grant number: 20/00088-9
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): March 01, 2020
• Effective date (End): February 28, 2022
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Cooperation agreement: MRC, UKRI ; Newton Fund, with FAPESP as a partner institution in Brazil
• Principal Investigator: Angela Kaysel Cruz
• Grantee: José Carlos Quilles Junior
• Home Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 18/14398-0 - UK:Brazil Joint Centre Partnership in Leishmaniasis (JCPiL), AP.TEM

Abstract

Control of gene expression in Leishmania occurs mainly at the post-transcriptional level. Central to the regulatory complexes are cis and trans-regulatory elements, mainly represented by mRNA elements (cis elements) and RNA binding proteins (RBPs), and may include hitherto unexplored content of noncoding transcripts (ncRNAs) recently detected in these parasites. We have previously demonstrated that ncRNAs appear to be a common feature of Leishmania transcripts, and approximately 300 putative ncRNAs were identified as differential expression (DE) transcripts. DE Leishmania braziliensis ncRNAs and RBPs interacting with them will be functionally investigated. State-of the-art technologies will be used for genome editing to generate knockout (KO) of ncRNAs DE in L. braziliensis and to endogenously tag these ncRNAs to evaluate phenotypic changes and identify interacting partners. The CRISPR/Cas9 system will be used for the KO of approximately one hundred small ncRNAs (transcripts of about 20-200 nucleotides) by associating this technology with a barcoding gene identification procedure. KO parasites for ncRNAs will be examined for their infection profiles capacity by in vitro infection assays. (AU)