title: Light-activated RNA interference in human embryonic stem cells.
abstract: We describe a near infrared (NIR) light-activated gene silencing method in undifferentiated human embryonic stem cell (hESC) using a plasmonic hollow gold nanoshell (HGN) as the siRNA carrier. Our modular biotin-streptavidin coupling strategy enables positively charged TAT-peptide to coat oligonucleotides-saturated nanoparticles as a stable colloid formation. TAT-peptide coated nanoparticles with dense siRNA loading show efficient penetration into a wide variety of hESC cell lines. The siRNA is freed from the nanoparticles and delivered to the cytosol by femtosecond pulses of NIR light with potentially exquisite spatial and temporal control. The effectiveness of this approach is shown by targeting GFP and Oct4 genes in undifferentiated hESC (H9). The accelerated expression of differentiation markers for all three germ layers resulting from Oct4 knockdown confirms that this method has no detectable adverse effects that limit the range of differentiation. This biocompatible and NIR laser-activated patterning method makes possible single cell resolution of siRNA delivery for diverse studies in stem cell biology, tissue engineering and regenerative medicine.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Barbara, CA 93106, United States.
authors.forename: Xiao
authors.initials: X
authors.lastname: Huang
authors.affiliation: Center for Stem Cell Biology and Engineering, University of California, Santa Barbara, CA 93106, United States.
authors.forename: Qirui
authors.initials: Q
authors.lastname: Hu
authors.affiliation: Cancer Research Center, Sanford-Burnham Medical Research Institute, La Jolla, CA 92037, United States.
authors.forename: Gary B
authors.initials: GB
authors.lastname: Braun
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Barbara, CA 93106, United States.
authors.forename: Alessia
authors.initials: A
authors.lastname: Pallaoro
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Barbara, CA 93106, United States.
authors.forename: Demosthenes P
authors.initials: DP
authors.lastname: Morales
authors.affiliation: Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, MN 55455, United States.
authors.forename: Joseph
authors.initials: J
authors.lastname: Zasadzinski
authors.affiliation: Center for Stem Cell Biology and Engineering, University of California, Santa Barbara, CA 93106, United States.
authors.forename: Dennis O
authors.initials: DO
authors.lastname: Clegg
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Santa Barbara, CA 93106, United States. Electronic address: reich@chem.ucsb.edu.
authors.forename: Norbert O
authors.initials: NO
authors.lastname: Reich
mesh_terms: D001710:Biotin / Q000737:chemistry; D002460:Cell Line; D015696:Gene Products, tat / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000235:genetics; D000066449:Human Embryonic Stem Cells / Q000378:metabolism*; D006801:Humans; D008027:Light; D053769:Nanocapsules / Q000737:chemistry*; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D010446:Peptide Fragments / Q000737:chemistry; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection
chemical_list: D015696:Gene Products, tat; D053769:Nanocapsules; D050814:Octamer Transcription Factor-3; D010446:Peptide Fragments; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D001710:Biotin; D006046:Gold
keywords: Differentiation; Hollow gold nanoshell; Human embryonic stem cells; Near-infrared light; RNA interference; TAT peptide