Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Department of Molecular Biology and Centre for Computational and Integrative Biology, Department of Genetics, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
Adv Mater. 2024 Oct;36(41):e2409356. doi: 10.1002/adma.202409356. Epub 2024 Aug 16.
Delivery of proteins in plant cells can facilitate the design of desired functions by modulation of biological processes and plant traits but is currently limited by narrow host range, tissue damage, and poor scalability. Physical barriers in plants, including cell walls and membranes, limit protein delivery to desired plant tissues. Herein, a cationic high aspect ratio polymeric nanocarriers (PNCs) platform is developed to enable efficient protein delivery to plants. The cationic nature of PNCs binds proteins through electrostatic. The ability to precisely design PNCs' size and aspect ratio allowed us to find a cutoff of ≈14 nm in the cell wall, below which cationic PNCs can autonomously overcome the barrier and carry their cargo into plant cells. To exploit these findings, a reduction-oxidation sensitive green fluorescent protein (roGFP) is deployed as a stress sensor protein cargo in a model plant Nicotiana benthamiana and common crop plants, including tomato and maize. In vivo imaging of PNC-roGFP enabled optical monitoring of plant response to wounding, biotic, and heat stressors. These results show that PNCs can be precisely designed below the size exclusion limit of cell walls to overcome current limitations in protein delivery to plants and facilitate species-independent plant engineering.
蛋白质在植物细胞中的递送可以通过调节生物过程和植物特性来促进期望功能的设计，但目前受到宿主范围狭窄、组织损伤和较差的可扩展性的限制。植物中的物理屏障，包括细胞壁和细胞膜，限制了蛋白质递送到期望的植物组织。本文开发了一种阳离子高纵横比聚合物纳米载体（PNC）平台，以实现蛋白质向植物的有效递送。PNC 的阳离子性质通过静电作用与蛋白质结合。精确设计 PNC 大小和纵横比的能力使我们能够找到细胞壁中的≈14nm 的截止值，低于该值，阳离子 PNC 可以自主克服障碍并将其货物带入植物细胞。为了利用这些发现，将氧化还原敏感的绿色荧光蛋白（roGFP）作为应激传感器蛋白货物部署在模式植物烟草和常见作物植物，包括番茄和玉米中。PNC-roGFP 的体内成像使我们能够光学监测植物对创伤、生物和热胁迫的反应。这些结果表明，PNC 可以精确设计成小于细胞壁的尺寸排除限制，以克服当前蛋白质递送到植物中的限制，并促进与物种无关的植物工程。
