Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA, 94720, USA.
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, 94720, USA.
Commun Biol. 2023 Aug 12;6(1):840. doi: 10.1038/s42003-023-05191-5.
Using a fluorescence complementation assay, Delivered Complementation in Planta (DCIP), we demonstrate cell-penetrating peptide-mediated cytosolic delivery of peptides and recombinant proteins in Nicotiana benthamiana. We show that DCIP enables quantitative measurement of protein delivery efficiency and enables functional screening of cell-penetrating peptides for in-planta protein delivery. Finally, we demonstrate that DCIP detects cell-penetrating peptide-mediated delivery of recombinantly expressed proteins such as mCherry and Lifeact into intact leaves. We also demonstrate delivery of a recombinant plant transcription factor, WUSCHEL (AtWUS), into N. benthamiana. RT-qPCR analysis of AtWUS delivery in Arabidopsis seedlings also suggests delivered WUS can recapitulate transcriptional changes induced by overexpression of AtWUS. Taken together, our findings demonstrate that DCIP offers a new and powerful tool for interrogating cytosolic delivery of proteins in plants and highlights future avenues for engineering plant physiology.
利用荧光互补测定法（Delivered Complementation in Planta，DCIP），我们证明了细胞穿透肽介导的肽和重组蛋白在本氏烟中的细胞质递送。我们表明，DCIP 能够定量测量蛋白递送效率，并能够对细胞穿透肽进行功能筛选，以实现植物体内蛋白递送。最后，我们证明了 DCIP 可以检测到重组表达的蛋白（如 mCherry 和 Lifeact）通过细胞穿透肽递送到完整叶片中。我们还证明了将重组植物转录因子 WUSCHEL（AtWUS）递送到本氏烟中。对拟南芥幼苗中 AtWUS 递送到 Arabidopsis seedlings 的 RT-qPCR 分析也表明，递送到的 WUS 可以重现由过表达 AtWUS 诱导的转录变化。总之，我们的发现表明，DCIP 为研究植物中蛋白的细胞质递送提供了一种新的强大工具，并为植物生理学的工程设计提供了未来的途径。
