Virus-induced CRISPR-Cas9 system improved resistance against tomato yellow leaf curl virus
- 15 April 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Molecular Biology Reports
- Vol. 47 (5), 3369-3376
- https://doi.org/10.1007/s11033-020-05409-3
Abstract
Plant viruses are the most significant factors associated with massive economical losses in agricultural industries worldwide. Accordingly, many studies are dedicated to making virus-resistant crop varieties each year due to the ever-changing nature of viruses. Recently genome engineering methods have been used to confer interference against eukaryotic viruses. Research results on genome editing technics, in particular, CRISPR-Cas9, promises a feasible solution to make virus-resistant crops. In this research, we explored the possibility of utilizing CRISPR-Cas9 to obtain TYLCV resistant tomato varieties. Moreover, to overcome any potential off-target effects of Cas9, we used an inducible promoter to initiate Cas9 activity in case of the virus attack. Cas9 vector was transformed by the rgsCaM promoter, known as an endogenous silencer of RNAi and overexpressed after a virus attack. The golden gate cloning method was applied to construct sgRNAs. Intergenic region and coat protein-coding sequences of TYLCV were used to design sgRNAs. Infiltrated sensitive Money Maker varieties analyzed by real-time PCR, showed a significant reduction or delayed accumulation of viral DNA compared to the control plants. This result demonstrates the efficiency of using an inducible promoter in CRISPR-Cas9 constructs.Keywords
Funding Information
- Ferdowsi University of Mashhad (3/41501)
- Biotechnology Development Council of the Islamic Republic of Iran (950709)
This publication has 26 references indexed in Scilit:
- CRISPR/Cas9 in Genome Editing and BeyondAnnual Review of Biochemistry, 2016
- Establishing a CRISPR–Cas-like immune system conferring DNA virus resistance in plantsNature Plants, 2015
- Conferring resistance to geminiviruses with the CRISPR–Cas prokaryotic immune systemNature Plants, 2015
- Management of Whitefly-Transmitted Viruses in Open-Field Production SystemsPublished by Elsevier BV ,2014
- The Tomato Yellow Leaf Curl Virus Resistance Genes Ty-1 and Ty-3 Are Allelic and Code for DFDGD-Class RNA–Dependent RNA PolymerasesPLoS Genetics, 2013
- Inhibition of Binding of Tomato Yellow Leaf Curl Virus Rep to its Replication Origin by Artificial Zinc-Finger ProteinMolecular Biotechnology, 2012
- The Spread of Tomato Yellow Leaf Curl Virus from the Middle East to the WorldPLoS Pathogens, 2010
- Use of Tomato yellow leaf curl virus (TYLCV) Rep Gene Sequences to Engineer TYLCV Resistance in TomatoPhytopathology®, 2004
- Tomato yellow leaf curl virus, the intracellular dynamics of a plant DNA virusMolecular Plant Pathology, 2002
- Tomato yellow leaf curl virus, an emerging virus complex causing epidemics worldwideVirus Research, 2000