Applications of multiplex genome editing in higher plants
Affiliation:
1. Department of Crop Production and Forestry Sciences, University of Lleida, Agrotecnio Center, Avda Alcalde Rovira Roure 191, 28049 Lleida, Spain;2. TRM Ltd., P.O. Box 493, Scarborough, YO11 9FJ, United Kingdom;3. ICREA, Catalan Institute for Research and Advanced Studies, Passeig Lluís Companys 23, 08010 Barcelona, Spain;1. School of Life Sciences, Central University of Gujarat, Gandhinagar, 382 030, Gujarat, India;2. Department of Botany, Patna Science College, Patna University, Patna, 800 005, Bihar, India;3. School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110 067, India;4. Crop Improvement Division, ICAR-Indian Institute of Vegetable Research, Varanasi, 221 305, Uttar Pradesh, India;1. Department of Plant Production and Forestry Science, School of Agrifood and Forestry Science and Engineering (ETSEA), University of Lleida-Agrotecnio Center, Lleida, Spain;2. Institute for Molecular Biotechnology, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany;3. TRM Ltd, PO Box 463, York YO11 9FJ, United Kingdom;4. Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstrasse 6, 52074 Aachen, Germany;5. ICREA, Catalan Institute for Research and Advanced Studies, Passeig Lluís Companys 23, 08010 Barcelona, Spain;1. Department of Pathology, Yale School of Medicine, New Haven, CT 06520, United States;2. Department of Pharmacology, Yale School of Medicine, New Haven, CT 06520, United States;1. National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan;2. Laboratory for Genome Engineering, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal, Saudi Arabia;1. Key Lab of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People’s Republic of China, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, 151 Malianwa North Road, Haidian District, Beijing 100193, China;2. Engineering Research Center of Chinese Medicine Resource, Ministry of Education, 151 Malianwa North Road, Haidian District, Beijing 100193, China
Abstract:
Multiplex genome editing in plants is typically carried out using the CRISPR/Cas9 system because it is more amenable to multiplexing than any other platform. A single gRNA can be used to target multiple alleles of the same gene (A1, A2) or homeoalleles from different ancestral genomes in a polyploid plant (A1, A2, B1, B2). A single gRNA can also be used to target multiple genes (X, Y, Z) if they contain a conserved region (C). Multiple gRNAs can be used to target unrelated sites in the same gene or in conserved sequences such as homeoalleles or members of the same gene family. Multiple gRNAs can also be used to target completely unrelated genes (D, E, F).
