CRISPR/Cas9 based targeted insertion of cystatins in a pre-characterized locus of sugarcane

• Grant number: 18/11544-5
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: December 10, 2018
• End date: December 09, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Flavio Henrique da Silva
• Grantee: Chakravarthi Mohan
• Supervisor: Fredy Altpeter
• Host Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil
• Institution abroad: University of Florida, Gainesville (UF), United States
• Associated to the scholarship: 15/10855-9 - Development of transgenic sugarcane more resistant to Sphenophorus levis, BP.PD

Abstract

Insect pests are among the most critical factors that compromise productivity of crops. Brazil has recently entered into a new area of crop protection by approving the world's first commercial GM sugarcane with a BT insect resistance gene. Sugarcane is one of the important cash crops cultivated worldwide for sugar and ethanol, with Brazil being the largest producer. Sphenophorus levis, popularly known as sugarcane weevil, is one of the critical pests of sugarcane leading to severe production losses in Brazil, especially in the state of São Paulo. Previous studies in our laboratory have reported that overexpression of cane cystatin I exhibited effective resistance to S. levis. However, the level and stability of transgene expression are critical for the predictable performance of a transgenic sugarcane cultivar. The standard transgenic technology leads to random insertions of transgenes associated with gene silencing and disruption of endogenous genes. Very few transgenic events display high level of transgene expression which depends on the genomic location where the transgene inserts. With the advent of genome editing technology, it is now feasible to target a desired gene at precise genomic location thereby eliminating the adverse position effects and leading to enhanced and predictable transgene performance. The current study aims to target cystatin gene in a pre-characterized, superior locus of sugarcane using CRISPR/Cas9 based genome editing tool. The proposed study will be carried out with the expertise of the research group of Prof. Dr. Fredy Altpeter, University of Florida, USA who have been recently successful in TALEN mediated targeted mutagenesis and CRISPR/Cas9 mediated homology directed gene targeting in sugarcane. (AU)

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)

EID, AYMAN; MOHAN, CHAKRAVARTHI; SANCHEZ, SARA; WANG, DUODUO; ALTPETER, FREDY. Multiallelic, Targeted Mutagenesis of Magnesium Chelatase With CRISPR/Cas9 Provides a Rapidly Scorable Phenotype in Highly Polyploid Sugarcane. FRONTIERS IN GENOME EDITING, v. 3, p. 12-pg., 2021-04-29. (18/11544-5)

ARENCIBIA, ARIEL D.; D'AFONSECA, VIVIAN; CHAKRAVARTHI, MOHAN; CASTIGLIONE, STEFANO. Learning from transgenics: Advanced gene editing technologies should also bridge the gap with traditional genetic selection. ELECTRONIC JOURNAL OF BIOTECHNOLOGY, v. 41, p. 22-29, SEP 2019. (18/11544-5)

MOHAN, CHAKRAVARTHI; EASTERLING, MONA; YAU, YUAN-YEU. ene Editing Technologies for Sugarcane Improvement: Opportunities and Limitation. SUGAR TECH, v. 24, n. 1, SI, OCT 2021. (15/10855-9, 18/11544-5)