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

这一行动为NSF国家植物基因组计划2020财年生物学博士后研究奖学金提供了资金。该奖学金支持在东道主实验室为该研究员制定的研究和培训计划，该研究员还提出了扩大生物学参与度的计划。乔恩·科迪的这项研究和培训计划的标题是“发育调节器介导的基因编辑：一种改善转基因植物可获得性和生产的方法”。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的改良，导致了农业生产力的提高，并推动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细胞并将其再生为完整的转基因植物，在许多作物物种中，这可能需要6-12个月的时间。目前的研究集中在优化和翻译最近的发现，证明在双子叶植物物种中，在没有组织培养的情况下，发育调节基因和激素生物合成基因的表达可以促进从头分生组织的产生。如果包括基因编辑试剂，就会产生转基因分生组织，将编辑传递给下一代。建立这一过程的简化方法可能会提高生产转基因和基因编辑双子叶作物的可及性和效率，从而广泛影响基因工程领域。在研究过程中，该研究员将接受植物生理学、分子生物学、基因工程、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培育转基因植物的过程，以及指导明尼苏达大学的本科生。该项目的主要目标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。初步工作表明，Cas9编辑试剂与Wus2和IPT编码序列共同传递可以促进编辑后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。本项目旨在提高发育调节剂转化技术的效率和翻译实用性-(1)优化：调控基因传递和Cas9介导的编辑的最佳策略是什么？(2)实用性：使用发育调节剂转化平台对本氏烟草进行同源定向修复是否可行？(3)翻译：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常规植物转化过程的困难，本工作的结果可能对植物生物学的研究产生广泛的影响。如果可能，研究人员可以使用发育调节剂介导的转化轻松地在具有农学意义的作物物种中创建基因编辑。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向公众和研究社区公布。这一奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.