Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
Molecular and Cellular Biology Graduate Program, University of Massachusetts, 230 Stockbridge Road, Amherst, Massachusetts 01003, United States.
Bioconjug Chem. 2021 May 19;32(5):891-896. doi: 10.1021/acs.bioconjchem.1c00103. Epub 2021 Apr 19.
Intracellular protein delivery is a transformative tool for biologics research and medicine. Delivery into the cytosol allows proteins to diffuse throughout the cell and access subcellular organelles. Inefficient delivery caused by endosomal entrapment is often misidentified as cytosolic delivery. This inaccuracy muddles what should be a key checkpoint in assessing delivery efficiency. Green fluorescent protein (GFP) is a robust cargo small enough to passively diffuse from the cytosol into the nucleus. Fluorescence of GFP in the nucleus is a direct readout for cytosolic access and effective delivery. Here, we highlight recent examples from the literature for the accurate assessment of cytosolic protein delivery using GFP fluorescence in the cytosol and nucleus.
细胞内蛋白质递送是生物医学研究和药物的一项变革性工具。递送到细胞质中可使蛋白质在整个细胞中扩散并进入亚细胞器。由于内体捕获而导致的低效递送通常被错误地识别为细胞质递送。这种不准确性使得评估递送效率的关键检查点变得模糊。绿色荧光蛋白（GFP）是一种足够小的货物蛋白，可以被动地从细胞质扩散到细胞核中。GFP 在细胞核中的荧光是细胞质进入和有效递送的直接读数。在这里，我们强调了文献中的最新示例，这些示例使用细胞质和细胞核中的 GFP 荧光准确评估了细胞质蛋白质递送。
