Alicai, Titus

Bart, Rebecca

Ingelbrecht, Ivan

Nusinow, Dmitri

Odipio, John

Taylor, Nigel

English

Ghent University Academic Bibliography

c Cassava (Manihot esculanta) is an important source of food, income and industrial raw materials. Rapid breeding of elite varieties to meet increasing demand is hampered by low rates of seed set, attributed to irregular flowering habits in many genotypes. The genetic basis of flowering in cassava is poorly understood. Studies in Arabidopsis and other species have identified key pathways and genes controlling enhancement and inhibition of flowering. CRISPR/CAS9 editing techniques offer novel, reliable alternatives for validation of gene function in plants. This study seeks to employ CRISPR/CAS9 technology for targeted modification of key flowering genes in cassava. As these pathways are also implicated in root tuberisation, information gained will increase understanding of both flowering and storage root development in cassava.  INTRODUCTION APPROACH ACKNOWLEDGEMENTS  Ceballos et al., 2015. DOI 10.1007/s00122-015-2555-4, Adeyemo et al. 2011. DOI 10.1007/s12042-011-9065-6; Pin P, Nilsson O. 2012. DOI: 10.1111/j.1365-3040.2012.02558.x; Navarro et al., 2011. Nature 478, 119–122; Nekrasov et al., 2013 DOI 10.1038/nbt.2655,  http://www.phytozome.net/)  REFERENCES CRISPR/CAS9 Genome Editing of Putative Cassava Flowering Genes  John Odipio1, 2, 3, Titus Alicai2, Dmitri Nusinow1, Rebecca Bart1,  Ivan Ingelbrecht3 and Nigel Taylor1 1Donald Danforth Plant Science Center, St. Louis, Missouri, USA, 2National Crops Resources Research Institute, Namulonge, Kampala, Uganda, 3VIB-Ghent University, Faculty of Science, Department of Plant Biotechnology and Bioinformatics, Ghent, Belgium   10 gRNAs designed and cloned for three targets   FEC produced as target tissue for gene editing and genetic transformation    ONGOING ACTIVITIES  ACCOMPLISHED ACTIVITIES  Design and screen new CRISPR/CAS9 constructs  Transform best constructs into cassava  Analyze and evaluate CAS9 edited plants for induced mutations and modified phenotypes  S  35 genes putatively associated with flowering identified from six gene families   1. Identify key genes & pathways controlling flowering and storage root development in cassava    2. Design and clone CRISPR/CAS9 constructs for modifying flowering phenotypes  3. Generate and analyze CAS9 edited cassava plants for induced mutations and modified flowering phenotypes    SPECIFIC OBJECTIVES  Validate CRISPR-gRNAs  constructs  Genome Sequence of Cassava Assemble CAS9 and gRNA into construct       CAS9 Vector   gRNA              Plasmid  Deliver CAS9-gRNA into FEC Select CAS9 edited events Evaluate CAS9 edited plants for induced mutations and modified phenotype Identify genes putatively associated with flowering Design gRNAs lacking  off-target potential Recover events with  induced mutations Analyze for induced mutations by restriction digestion & sequencing  

CRISPR/CAS9 genome editing of putative cassava flowering genes

https://biblio.ugent.be/publication/8550236/file/8550238.pdf

CRISPR/CAS9 genome editing of putative cassava flowering genes

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Ghent University Academic Bibliography