Development of an Odorant Co-­receptor Mutant Strain of the New World Screwworm Fly using the CRISPR/Cas9 system: A Functional Investigation on the Evolution of Olfactory Perception in Blowflies.

• Grant number: 17/05432-7
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: May 23, 2017
• End date: May 22, 2018
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Ana Maria Lima de Azeredo-Espin
• Grantee: Daniel Fernando Paulo
• Supervisor: Owen William Mcmillan
• Host Institution: Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Institution abroad: Smithsonian Institution, Panamá, Panama
• Associated to the scholarship: 14/15042-3 - Expression profile analysis of microRNAs during different development stages of the New World Screwworm fly Cochliomyia hominivorax (Diptera: Calliphoridae), BP.DR

Abstract

Identifying genomic regions that underlie the adaptive radiation of insects is a major goal in modern evolutionary biology. The emergence of obligatory ectoparasitism in blowflies has been the subject for scientific debate for over a half-century, creating the need for studies regarding how genomic architecture is linked to adaptive ecological transitions. Previous studies suggested that this strategy arose twice within blowflies' lineage, presumable to avoid resource competition. However, the molecular bases underlying the transition of a free-living to an ectoparasitic habit in blowflies remain unknown. We hypothesize that olfaction might have played a critical role in this adaptation. To begin testing this, this project propose to functionally characterize the olfactory co-receptor Orco in the New World Screwworm fly (NWS), Cochiomyia hominivorax, by using the powerful genome editing CRISPR/Cas9 system to knockout this gene. Orco encodes a co-receptor that dimerizes with all Odorant-selective Receptors (ORs) and is required for the OR-perception pathway. After knocking the gene out in NWS, a stable Orco mutant strain will be developed, which functionally lacks the entire ORs family. This project focus on the establishment of the CRISPR system in NWS, but it will also provide important information about the odors associated with the ectoparasitism in blowflies by generating the first profile of responses mediated by olfactory receptors in C. hominivorax.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

PAULO, DANIEL F.; WILLIAMSON, MEGAN E.; ARP, ALEX P.; LI, FANG; SAGEL, AGUSTIN; SKODA, STEVEN R.; SANCHEZ-GALLEGO, JOEL; VASQUEZ, MARIO; QUINTERO, GLADYS; PEREZ DE LEON, ADALBERTO A.; et al. Specific Gene Disruption in the Major Livestock Pests Cochliomyia hominivorax and Lucilia cuprina Using CRISPR/Cas9. G3-GENES, GENOMES, GENETICS, v. 9, n. 9, p. 3045-3055, SEP 2019. (17/05432-7)

PAULO, DANIEL F.; JUNQUEIRA, ANA C. M.; ARP, ALEX P.; VIEIRA, ANDRE S.; CEBALLOS, JORGE; SKODA, STEVEN R.; PEREZ-DE-LEON, ADALBERTO A.; SAGEL, AGUSTIN; MCMILLAN, WILLIAM O.; SCOTT, MAXWELL J.; et al. Disruption of the odorant coreceptor Orco impairs foraging and host finding behaviors in the New World screwworm fly. SCIENTIFIC REPORTS, v. 11, n. 1, MAY 31 2021. (14/15042-3, 15/02079-9, 17/05432-7)