RNA delivery by heat shock protein-70 into mammalian cells: a preliminary study.
Heat shock proteins (Hsps) are ubiquitous molecular chaperones with indispensable roles in assisting protein folding and giving protection from proteotoxic environmental harm. Members of the 70-kDa heat shock protein family have been demonstrated to recognize and bind with distinguished RNA sequences, which function as determinants of eukaryotic mRNA stability. We have earlier identified the molecular domains involved in RNA-binding and characterized in detail the specificity, affinity and some regulatory aspects of this molecular interaction using various deletion mutants and homologues of Hsp70. We have shown that wild type, but not any of the tested truncated mutants of Hsp70, is efficiently taken up by P388 mouse macrophage cells. Here we addressed the question of whether Hsp70 is capable of delivering bound RNA into mammalian cells. Employing fluorescence and confocal microscopy, we demonstrated that full length Hsp70 facilitates the uptake of RNA molecules into the cytoplasm of mammalian cells. We propose that further optimization of this system might enable the development of a valuable tool to deliver RNA molecules, such as siRNA, dsRNA or other regulatory RNA sequences to probe or influence various regulatory processes in eukaryotic cells.
INTERCELL AG, Campus Vienna Biocenter, 1030 Vienna, Austria. tamas.henics@univie.ac.at
Tamás
T
Henics
Christine
C
Zimmer
D000818:Animals; D002478:Cells, Cultured; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D018832:Molecular Chaperones / Q000378:metabolism; D033704:Nucleotide Transport Proteins / Q000378:metabolism*; D010865:Pilot Projects; D011485:Protein Binding; D012313:RNA / Q000378:metabolism*; D015347:RNA Probes
D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones; D033704:Nucleotide Transport Proteins; D015347:RNA Probes; D012313:RNA