Generation of T. cruzi (Y strain) knockout (KO) lineages for IP6K, TERT and IP6K+TERT using the CRISPR-Cas9 approach

• Grant number: 23/09435-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2023
• End date: July 31, 2024
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Marcelo Santos da Silva
• Grantee: Giovana Zukowski Luduvice
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 19/10753-2 - Investigation on the role of inositol pyrophosphates (PP-IPs) in DNA repair pathways and telomere dynamics using trypanosomatids as a model, AP.JP

Abstract

In model eukaryotes, inositol pyrophosphates (PP-IPs) - mainly IP7 and IP8 - are involved in a wide range of cellular processes, such as regulation of telomere length and homologous recombination (HR). However, the mechanism of action of these metabolites is still not well established. The main PP-IPs involved in these processes are IP7 and IP8, which are synthesized by pathways involving the participation of IP6K and PP-IP5K kinases. Trypanosomatid parasites do not have orthologous genes for PPIP5K, that is, they possibly do not synthesize IP8, which makes them excellent models for the study of IP7. Thus, this scientific initiation project consists of generating strains of T. cruzi (Y strain) KO for IP6K (IP6K-/- or IP6K-/+), telomerase (TERT-/-) and double-KO (IP6K-/- + TERT-/-), which will facilitate the subsequent investigation of the role of IP7 in DNA repair pathways and telomere homeostasis. These T. cruzi strains will be essential to answer fundamental questions raised by the main project to which this grant is linked.