An Agrobacterium ‐delivered CRISPR /Cas9 system for high‐frequency targeted mutagenesis in maize
2016; Wiley; Volume: 15; Issue: 2 Linguagem: Inglês
10.1111/pbi.12611
ISSN1467-7652
AutoresSi Nian Char, Anjanasree K. Neelakandan, Hartinio N. Nahampun, Bronwyn Frame, Marcy Main, Martin H. Spalding, Philip W. Becraft, Blake C. Meyers, Virginia Walbot, Kan Wang, Bing Yang,
Tópico(s)Plant tissue culture and regeneration
ResumoCRISPR/Cas9 is a powerful genome editing tool in many organisms, including a number of monocots and dicots. Although the design and application of CRISPR/Cas9 is simpler compared to other nuclease-based genome editing tools, optimization requires the consideration of the DNA delivery and tissue regeneration methods for a particular species to achieve accuracy and efficiency. Here, we describe a public sector system, ISU Maize CRISPR, utilizing Agrobacterium-delivered CRISPR/Cas9 for high-frequency targeted mutagenesis in maize. This system consists of an Escherichia coli cloning vector and an Agrobacterium binary vector. It can be used to clone up to four guide RNAs for single or multiplex gene targeting. We evaluated this system for its mutagenesis frequency and heritability using four maize genes in two duplicated pairs: Argonaute 18 (ZmAgo18a and ZmAgo18b) and dihydroflavonol 4-reductase or anthocyaninless genes (a1 and a4). T
Referência(s)