Identification of host cell genes that affect mRNA retention and HIV Rev-RRE function

• Grant number: 19/09887-4
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• Start date: July 08, 2019
• End date: July 07, 2020
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Edison Luiz Durigon
• Grantee: Flávio da Silva Mesquita
• Supervisor: Marie-Louise Hammarskjold
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: University of Virginia (UVa), United States
• Associated to the scholarship: 18/10913-7 - Development of a new in vitro inhibition strategy for Zika Virus and Influenza Virus using CRISPR-Cas13 RNA-editing technique as an alternative treatment, BP.DD

Abstract

The project will involve the use of several molecular biological methods and bioinformatic tools to study HIV. In Aim #1, long read sequencing methods using nanopore technology will be developed to study different splicing isoforms expressed from a novel HIV vector. Aim #2 will serve to identify host cell genes involved in nuclear retention of HIV mRNAs, as well as host genes that positively or negatively affect Rev function. Rev is a viral nuclear export protein that is known to interact with the host cell protein Crm1, which serves as an export protein for proteins as well as mRNAs. Although it has been known for many years that Rev promotes the nucleocytoplasmic export of HIV mRNAs, many aspects of Rev regulation remain unknown. Specifically, in Aim #1, we will sequence and analyze all the different isoforms of HIV mRNAs that are expressed from a novel HIV- based vector. In Aim #2 we will use CRISPR/Cas9 Lentiviral vector libraries in conjunction with the new HIV vector system to identify host cell genes that are involved in nuclear retention of HIV mRNAs with retained introns, as well as novel host genes that are involved in Rev/RRE function. (AU)