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Different cell-penetrating peptides (CPPs) for delivery of green fluorescent protein (GFP) into mammalian cells

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posted on 2024-09-18, 10:34 authored by Sanjay PatelSanjay Patel, Edward Sayers, Lin He, Rohan Narayan, Thomas Williams, Rudolf AllemannRudolf Allemann, Louis LukLouis Luk, Arwyn JonesArwyn Jones, Yu-Hsuan TsaiYu-Hsuan Tsai

We use green fluorescent protein (GFP) as a model cargo to compare the delivery capability of five CPP sequences (Penetratin, R8, TAT, Transportan, Xentry) and their derivatives (cyclic R8, cyclic TAT, TAT-HA) in four cell lines (HeLa, HEK, 10T1/2, HepG2). 

For confocal microscopy experiments, HeLa cells were incubated with 10 µM of the designated eGFP-CPP fusion proteins at 37 °C for 1 h before imaging. Nucleus were stained with Hoechst which shows blue fluorescence. Biological duplicates of confocal microscopy results (*.lif) are available here. 

For flow cytometry experiments, cells were treated with 10 µM of eGFP-CPP fusion proteins at 37 °C for 1 h before analysis. Biological triplicates of flow cytometry results (*.fcs) are available here. 

Funding

Traceless, non-invasive and spatiotemporal control of protein activity in cells

Biotechnology and Biological Sciences Research Council

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Cleavable linkers for traceless delivery of therapeutics into cells (2016-09-01 - 2017-03-31); Tsai, Yu-Hsuan. Funder: Engineering and Physical Sciences Research Council

History

Specialist software required to view data files

ImageJ and ProSort were used to view .lif and .fcs files, respectively

Data-collection start date

2016-11-01

Data-collection end date

2018-10-31

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    School of Chemistry

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