Photosensitizing Carrier Proteins for Photoinducible RNA Interference

Matsushita-Ishiodori, Yuka; Kuwabara, Rina; Sakakoshi, Hiroyuki; Endoh, Tamaki; Ohtsuki, Takashi

doi:10.1021/bc200095a

Cited by 23 publications

(17 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast to the caged DNA and RNA nucleobases discussed below, one approach pioneered by the Ohtsuki lab utilizes a protein carrier to deliver RNA cargo (i.e., siRNA or shRNA) into cells through the conjugation of photosensitizer dyes to cell-permeable RNA-binding proteins (RBPs). [128] In the absence of light, a significant portion of the RNA/RBP complex remains trapped in endosomes and is eventually degraded. However, illumination with light affords improved endosomal escape and subsequent release of the RNA cargo into the cytosol.…”

Section: Optical Control Of Proteins and Peptidesmentioning

confidence: 99%

Optochemical Control of Biological Processes in Cells and Animals

et al. 2018

View full text Add to dashboard Cite

Biological processes are naturally regulated with high spatial and temporal control, as is perhaps most evident in metazoan embryogenesis. Chemical tools have been extensively utilized in cell and developmental biology to investigate cellular processes, and conditional control methods have expanded applications of these technologies toward resolving complex biological questions. Light represents an excellent external trigger since it can be controlled with very high spatial and temporal precision. To this end, several optically regulated tools have been developed and applied to living systems. In this review we discuss recent developments of optochemical tools, including small molecules, peptides, proteins, and nucleic acids that can be irreversibly or reversibly controlled through light irradiation, with a focus on applications in cells and animals.

show abstract

Section: Optical Control Of Proteins and Peptidesmentioning

confidence: 99%

Optochemical Control of Biological Processes in Cells and Animals

et al. 2018

View full text Add to dashboard Cite

show abstract

“…This carrier molecule can bind to an RNA and deliver it into a cell by the PCI strategy, whereby the carrier/RNA complexes are first entrapped within endosomes, and then photo-dependently escape the endosomes. The photosensitizing RNA carrier molecule consists of a photosensitizer and TatU1A, a fusion protein of HIV TAT-derived cell-penetrating peptide (CPP) and human U1A-derived RNA-binding protein 13 14 15 . An advantage to the use of a photosensitizer attached to the TatU1A protein for studying the PCI mechanism is that the localization of photosensitizers (~1 kDa) is strongly affected by the larger protein moiety (~16 kDa).…”

mentioning

confidence: 99%

The molecular mechanism of photochemical internalization of cell penetrating peptide-cargo-photosensitizer conjugates

Ohtsuki

Miki

Kobayashi³

et al. 2015

Sci Rep

Self Cite

View full text Add to dashboard Cite

In many drug delivery strategies, an inefficient transfer of macromolecules such as proteins and nucleic acids to the cytosol often occurs because of their endosomal entrapment. One of the methods to overcome this problem is photochemical internalization, which is achieved using a photosensitizer and light to facilitate the endosomal escape of the macromolecule. In this study, we examined the molecular mechanism of photochemical internalization of cell penetrating peptide-cargo (macromolecule)-photosensitizer conjugates. We measured the photophysical properties of eight dyes (photosensitizer candidates) and determined the respective endosomal escape efficiencies using these dyes. Correlation plots between these factors indicated that the photogenerated 1O2 molecules from photosensitizers were highly related to the endosomal escape efficiencies. The contribution of 1O2 was confirmed using 1O2 quenchers. In addition, time-lapse fluorescence imaging showed that the photoinduced endosomal escape occurred at a few seconds to a few minutes after irradiation (much longer than 1O2 lifetime), and that the pH increased in the endosome prior to the endosomal escape of the macromolecule.

show abstract

“…It is particularly easy to use already available Fucci-expressing cell lines, such as NMuMG/Fucci and COS/ Fucci in the RIKEN cell bank. Moreover, the system may also potentially be applied for studying the cell cycle dependence of RNA functions as a photoinduced cytoplasmic RNA delivery system has been developed 27,41,42 .…”

Section: Resultsmentioning

confidence: 99%

Cell cycle dependence of apoptosis photo-triggered using peptide-photosensitizer conjugate

Kim

Watanabe

Kitamatsu

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

Investigation of the relevance between cell cycle status and the bioactivity of exogenously delivered biomacromolecules is hindered by their time-consuming cell internalization and the cytotoxicity of transfection methods. In this study, we addressed these problems by utilizing the photochemical internalization (PCI) method using a peptide/protein-photosensitizer conjugate, which enables immediate cytoplasmic internalization of the bioactive peptides/proteins in a light-dependent manner with low cytotoxicity. To identify the cell-cycle dependent apoptosis, a TatBim peptide-photosensitizer conjugate (TatBim-PS) with apoptotic activity was photo-dependently internalized into HeLa cells expressing a fluorescent ubiquitination-based cell cycle indicator (Fucci2). Upon irradiation, cytoplasmic TatBim-PS internalization exceeded 95% for all cells classified in the G1, S, and G2/M cell cycle phases with no significant differences between groups. TatBim-PS-mediated apoptosis was more efficiently triggered by photoirradiation in the G1/S transition than in the G1 and S/G2/M phases, suggesting high sensitivity of the former phase to Bim-induced apoptosis. Thus, the cell cycle dependence of Bim peptide-induced apoptosis was successfully investigated using Fucci2 indicator and the PCI method. Since PCI-mediated cytoplasmic internalization of peptides is rapid and does not span multiple cell cycle phases, the Fucci-PCI method constitutes a promising tool for analyzing the cell cycle dependence of peptides/protein functions.

show abstract

Photosensitizing Carrier Proteins for Photoinducible RNA Interference

Cited by 23 publications

References 22 publications

Optochemical Control of Biological Processes in Cells and Animals

Optochemical Control of Biological Processes in Cells and Animals

The molecular mechanism of photochemical internalization of cell penetrating peptide-cargo-photosensitizer conjugates

Cell cycle dependence of apoptosis photo-triggered using peptide-photosensitizer conjugate

Contact Info

Product

Resources

About