NSF Postdoctoral Fellowship in Biology FY 2020: Developmental Regulator-Mediated Gene Editing: A Method to Improve Accessibility and Production of Transgenic Plants

该行动资助了2020财年NSF国家植物基因组计划生物学博士后研究奖学金。该研究金支持研究员在东道实验室的研究和培训计划，研究员还提出了扩大生物学参与的计划。该奖学金的研究和培训计划的标题是“发育调节因子介导的基因编辑：一种改善转基因植物可及性和生产的方法”。该奖学金的主办机构是明尼苏达大学，赞助科学家是丹尼尔·沃伊塔斯博士。在过去的30年里，传统的基因工程方法通过扩大可用性状加速了植物改良，导致农业生产力的提高，并促进了新的生物机制的发现。然而，这一过程需要使用组织培养技术来选择编辑过的细胞并将其再生为完整的转基因植物，这在许多作物物种中可能需要6 - 12个月。目前的研究重点是优化和翻译最近的研究结果，表明发育调节因子和激素生物合成基因的表达可以促进双子叶植物物种在没有组织培养的情况下从头分生组织的产生。如果包括基因编辑试剂，则产生转基因分生组织，将编辑传递给下一代。建立用于该过程的简化方法可以通过可能增加生产转基因和基因编辑的双子叶作物植物的可及性和效率来广泛影响基因工程领域。在这项研究过程中，研究员将接受植物生理学、分子生物学、遗传工程、基因组学、表型组学和植物转化等学科的培训。更广泛的影响包括在科学会议上传播研究结果，制作视频教程，详细介绍使用发育调节剂创造转基因植物的过程，并指导明尼苏达大学的本科生。该项目的主要目的是研究发育调节剂介导的异位分生组织形成的可能用途，以在没有组织培养的情况下为作物物种创造一种有效和可获得的基因工程方法。初步工作已经证明，Cas9编辑试剂与Wus 2和ipt编码序列一起的共递送可以促进编辑的从头分生组织的形成。然而，这份初步报告是一个概念证明，需要优化。本项目的主要目的是提高发育调节基因转化技术的效率和转化效用-（1）优化：调节基因递送和Cas9介导的编辑的最佳策略是什么？(2)实用性：在本氏烟草中使用发育调节因子转化平台进行同源定向修复可行吗？(3)已建立的发育调节剂转化管道能否用于农艺作物物种，如番茄和马铃薯？由于基因工程的广泛应用和传统植物转化过程的困难，本工作的结果可能对植物生物学研究产生广泛的影响。如果可能的话，研究人员可以使用发育调节剂介导的转化来轻松地在农艺学上重要的作物物种中进行基因编辑。这项工作的结果和重要观察结果将通过出版物和基于视频的协议提供给公众和研究界。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识价值和更广泛的影响审查标准进行评估来支持。

This action funds an NSF National Plant Genome Initiative Postdoctoral Research Fellowship in Biology for FY 2020. The fellowship supports a research and training plan in a host laboratory for the Fellow who also presents a plan to broaden participation in biology. The title of the research and training plan for this fellowship to Jon Cody is "Developmental Regulator-Mediated Gene Editing: A Method to Improve Accessibility and Production of Transgenic Plants". The host institution for the fellowship is the University of Minnesota and the sponsoring scientist is Dr. Daniel Voytas.For the past 30 years, conventional genetic engineering methods have accelerated plant improvement through the expansion of available traits, leading to an increase in agricultural productivity and fueling the discovery of new biological mechanisms. This process, however, requires the use of tissue culture techniques to select and regenerate edited cells into whole transgenic plants, which can take 6 – 12 months in many crop species. The presented research focuses on optimization and translation of recent findings that demonstrate expression of developmental regulators and hormone biosynthetic genes can promote the production of de novo meristems in the absence of tissue culture in dicot plant species. If gene editing reagents are included, transgenic meristems are produced that transmit edits to the next generation. Establishment of streamlined methods for this process could broadly impact the field of genetic engineering by possibly increasing accessibility and efficiency of producing transgenic and gene edited dicot crop plants. During the course of this study the fellow will be trained in the disciplines of plant physiology, molecular biology, genetic engineering, genomics, phenomics and plant transformation. Broader impacts include the dissemination of findings at scientific conferences, production of video tutorials detailing the process of creating transgenic plants using developmental regulators and mentoring undergraduate students at the University of Minnesota. The primary objective of the project is to investigate the possible utility of developmental regulator-mediated ectopic meristem formation to create an efficient and accessible genetic engineering method for crop species in the absence of tissue culture. Preliminary work has demonstrated codelivery of Cas9 editing reagents together with Wus2 and ipt coding sequences can promote formation of edited de novo meristems. However, this initial report was a proof of concept and requires optimization. This project focuses on the following aims to improve efficiency and translate utility of developmental regulator transformation technology - (1) Optimization: What is the best strategy for regulator gene delivery and Cas9-mediated editing? (2) Utility: Is homology directed repair in Nicotiana benthamiana using a developmental regulator transformation platform feasible? (3) Translation: Can the established developmental regulator transformation pipeline be utilized in agronomic crop species, such as tomato and potato? Due to the widespread application of genetic engineering and the difficulty of the conventional plant transformation process, the results from the present work could have a broad impact on research in plant biology. If possible, developmental regulator-mediated transformation could be used by researchers to easily create gene edits in agronomically significant crop species. Results and important observations from this work will be available to the public and research community through publications and video-based protocols.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.