Photo inducible RNA interference using cell permeable protein carrier.
In this study, we constructed fluorescently labelled protein carrier constructed from cell-penetrating peptide (CPP) and RNA binding domain (RBD) for intracellular shRNA or siRNA delivery. The protein carrier specifically bound to an RNA cargo containing short extended sequence tag for protein binding, and internalized into CHO cells together with the RNA. Although the internalized protein/RNA complexes showed cytoplasmic punctuate distributions, they widely spread into cytosol by photo irradiation. Moreover, redistributed RNA induced gene silencing only within the photo irradiated area.
Endoh
Tamaki
T
Department of Bioscience and Biotechnology, Okayama-University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
Sisido
Masahiko
M
Ohtsuki
Takashi
T
D000818:Animals; D016466:CHO Cells; D002463:Cell Membrane Permeability; D006224:Cricetinae; D003412:Cricetulus; D011992:Endosomes / Q000378:metabolism; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000528:radiation effects; D011806:Quinolinium Compounds; D034622:RNA Interference; D022661:RNA, Untranslated / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000737:chemistry / Q000378:metabolism; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry / Q000378:metabolism*
C481052:Alexa fluor 546; D005456:Fluorescent Dyes; D010455:Peptides; D011806:Quinolinium Compounds; D022661:RNA, Untranslated; D016601:RNA-Binding Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear; C075339:U1A protein; D054322:tat Gene Products, Human Immunodeficiency Virus