PASTE: a high-throughput method for large DNA insertions
Institution:
1. Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Constituent College of Pakistan Institute of Engineering and Applied Sciences, Jhang Road, Faisalabad, Pakistan;2. Department of Biological Sciences, University of Sialkot, Sialkot, Pakistan;3. International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan;1. The James Hutton Institute, Invergowrie, Dundee DD2 5DA, UK;2. Innovation Centre for Organic Farming, Agro Food Park 26, 8200 Aarhus N., Denmark;3. Agronomy Institute, Orkney College, University of the Highlands and Islands, Orkney, UK;4. Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, Thorvaldsensvej 40, 1879 Frederiksberg C., Denmark;1. National Institute of Science and Technology on Plant Physiology under Stress Conditions, Departamento de Biologia Vegetal, Universidade Federal de Viçosa, 36570-900 Viçosa, MG, Brazil;2. Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476 Potsdam-Golm, Germany;1. National Key Laboratory for Crop Genetics and Germplasm Enhancement and Utilization, Collaborative Innovation Center for Modern Crop Production Co-Sponsored by Province and Ministry (CIC-MCP), Nanjing Agricultural University, No.1 Weigang, Nanjing, Jiangsu 210095, China;1. Department of Plant Protection Biology, Swedish University of Agricultural Sciences, 23422 Alnarp, Sweden;2. Department of Ecology, Swedish University of Agricultural Sciences, 75007 Uppsala, Sweden;3. Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences, 75007 Uppsala, Sweden;4. Department of Molecular Sciences, Swedish University of Agricultural Sciences, 75007 Uppsala, Sweden;1. Institute of Molecular Bioscience, The University of Queensland, Brisbane, QLD 4072, Australia;2. School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia;3. Treble Cone Advisory Brisbane Qld, Suite 75, 12 Welsby Street, New Farm, QLD 4005, Australia;4. Centre for Policy Futures, Faculty of Humanities and Social Sciences, The University of Queensland, Brisbane, QLD 4072, Australia;5. Queensland Alliance for Agriculture and Food innovation, The University of Queensland, Brisbane, QLD 4072, Australia
Abstract:
Prime editing (PE) enables precise genome editing at targeted locus without inducing double-stranded breaks (DSBs). Despite its precision, PE lacks the tendency to integrate large DNA fragments into the genome. Recently, Yarnall et al. reported clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 and an integrase-based system that conducts targeted integration of large DNA sequences (~36 kb) into the genome more efficiently.
