Generation of L. braziliensis knockout (KO) lineages for IP6K, TERT and IP6K+TERT using CRISPR-Cas9 approach

• Grant number: 20/16481-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2021
• End date: February 28, 2022
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Marcelo Santos da Silva
• Grantee: Vitor Luiz da Silva
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , 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) are involved in a wide range of processes, such as telomere lengthening and homologous recombination (HR). However, the PP-IPs target-proteins, as well as their action mechanisms remains unknown. The main PP-IPs involved in these processes are PP-IP4, IP7 and IP8, which are synthesized by pathways involving the participation of IP6K and PP-IP5K kinases. Trypanosomatid parasites do not have ortholog genes for PP-IP5K, i.e., they do not synthesize IP8, which make them excellent models for the study of PPIP4 e IP7. Thus, this research plan consists in generates L. braziliensis lineages knockout for IP6K (IP6K-/-), telomerase (TERT-/-), and double-KO (IP6K-/- + TERT-/-), which will allow a subsequent investigation of the role of PP-IP4 and IP7 in DNA repair and telomere homeostasis pathways. These L. braziliensis lineages will be essential to answering fundamental questions raised by the main project.