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Cody, J. P.

Publications and source records attributed to Cody, J. P..

2 recordsLinked to original sources

Defining the Parameters to Improve Plant Regeneration with Developmental Regulators

Tissue culture methods which serve as the standard to regenerate modified plants are challenging and have limited the capacity to engineer new accessions. To improve upon these techniques, genome modifying reagents have been combined with developmental regulators to create gene edited plant tissues in both monocot and eudicot species. Co-culturing seedlings with Agrobacterium strains encoding developmental regulatory genes has proved to be effective at producing de novo meristems in multiple eudicot species. In order to see that this technology scales well beyond proof of concept experiments, various parameters were tested for refinement. Improvements have been observed at the key stages of growth induction and progression to shooting by manipulating the vector design, developmental regulator choice, Agrobacterium strain selection, and regulatory gene removal systems. Having defined these parameters as viable optimization points, the avenues to apply developmental regulators for plant regeneration in more diverse species have become more feasible.

plant biology↗

An Extensible Vector Toolkit and Parts Library for Advanced Engineering of Plant Genomes

Plant biotechnology is rife with new advances in transformation and genome engineering techniques. A common requirement for delivery and coordinated expression in plant cells, however, places the design and assembly of transformation constructs at a crucial juncture as desired reagent suites grow more complex. Modular cloning principles have simplified some aspects of vector design, yet many important components remain unavailable or poorly adapted for rapid implementation in biotechnology research. Here, we describe a universal Golden Gate cloning toolkit for vector construction. The toolkit chassis is compatible with the widely accepted Phytobrick standard for genetic parts, and supports assembly of arbitrarily complex T-DNAs through improved capacity, positional flexibility, and extensibility in comparison to extant kits. We also provision a substantial library of newly adapted Phytobricks, including regulatory elements for monocot and dicot gene expression, and coding sequences for genes of interest such as reporters, developmental regulators, and site-specific recombinases. Finally, we use a series of dual luciferase assays to measure contributions to expression from promoters, terminators, and from cross-cassette interactions attributable to enhancer elements in certain promoters. Taken together, these publicly available cloning resources can greatly accelerate the testing and deployment of new tools for plant engineering.

plant biology↗