Light-activated RNA interference in human embryonic stem cells

Xiao Huang, Qirui Hu, Gary B. Braun, Alessia Pallaoro, Demosthenes P. Morales, Joseph Zasadzinski, Dennis O. Clegg, Norbert O. Reich

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)70-79
Number of pages10
JournalBiomaterials
Volume63
DOIs
StatePublished - Sep 1 2015

Bibliographical note

Funding Information:
This work was supported by National Institutes of Health (NIH) grant R01 EB012637 and California Institute for Regenerative Medicine (CIRM) grant TG2-01151 . X.H. and Q.H. are CIRM fellows. The authors thank Mary Raven for help with dark-field microscopy and confocal microscopy, supported by NIH grant S10OD010610-01A1 . We also wish to acknowledge Sherry Hikita, Michelle Maloney, and Cassidy Hinman at the UCSB Laboratory for Stem Cell Biology and Engineering, supported by CIRM grant CL1-00521 . X.H. acknowledges support from Chinese Scholarship Council (CSC) file number 2011674001. G.B.B. acknowledges support from the NIH ( R01 CA 152327 , T32 CA 121949 ). Q.H. and D.O.C. acknowledge the Garland Initiative for Vision, CIRM grants DR1-0144, FA1-00616, the Foundation Fighting Blindness Wynn-Gund Translational Research Acceleration Program, and the UC Santa Barbara Institute for Collaborative Biotechnologies through grant W911NF-09-0001 from the U.S. Army Research Office. The content of the information does not necessarily reflect the position or the policy of the government, and no official endorsement should be inferred.

Publisher Copyright:
© 2015.

Keywords

  • Differentiation
  • Hollow gold nanoshell
  • Human embryonic stem cells
  • Near-infrared light
  • RNA interference
  • TAT peptide

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