3 research outputs found
Inducing Stem Cell Myogenesis Using NanoScript
Transcription factors (TFs) are multidomain proteins that play a critical role in orchestrating stem cell differentiation, but several limitations hinder the full potential of TF-based gene regulation. Here we report a unique strategy to emulate TFs and differentiate stem cells in a nonviral approach using an artificial, nanoparticle-based transcription factor called NanoScript. The NanoScript platform consists of a gold nanoparticle functionalized with small molecules that mimic the various domains of TFs. As a result, NanoScript mimics the function and structure of TF proteins. Specifically, NanoScript was designed to regulate muscle cell differentiation by targeting myogenic regulatory factors (MRFs), which play an important role in inducing myogenesis. This NanoScript-MRF is stable in physiological environments, localizes within the nucleus, induces differentiation of adipose-derived mesenchymal stem cells into mature muscle cells in 7 days, and is naturally excreted from induced muscle cells. As such, NanoScript represents a safe and powerful tool for applications requiring gene manipulation
NanoScript: A Nanoparticle-Based Artificial Transcription Factor for Effective Gene Regulation
Transcription factor (TF) proteins are master regulators of transcriptional activity and gene expression. TF-based gene regulation is a promising approach for many biological applications; however, several limitations hinder the full potential of TFs. Herein, we developed an artificial, nanoparticle-based transcription factor, termed NanoScript, which is designed to mimic the structure and function of TFs. NanoScript was constructed by tethering functional peptides and small molecules called synthetic transcription factors, which mimic the individual TF domains, onto gold nanoparticles. We demonstrate that NanoScript localizes within the nucleus and initiates transcription of a reporter plasmid by over 15-fold. Moreover, NanoScript can effectively transcribe targeted genes on endogenous DNA in a nonviral manner. Because NanoScript is a functional replica of TF proteins and a tunable gene-regulating platform, it has great potential for various stem cell applications
Integrating Epigenetic Modulators into NanoScript for Enhanced Chondrogenesis of Stem Cells
<i>N</i>-(4-Chloro-3-(trifluoroÂmethyl)Âphenyl)-2-ethoxyÂbenzamide
(CTB) is a small molecule that functions by altering the chromatin
architecture to modulate gene expression. We report a new CTB derivative
with increased solubility and demonstrate CTB’s functionality
by conjugating it on the recently established NanoScript platform
to enhance gene expression and induce stem cell differentiation. NanoScript
is a nanoparticle-based artificial transcription factor that emulates
the structure and function of transcription factor proteins (TFs)
to effectively regulate endogenous gene expression. Modifying NanoScript
with CTB will more closely replicate the TF structure and enhance
CTB functionality and gene expression. To this end, we first conjugated
CTB onto NanoScript and initiated a time-dependent increase in histone
acetylÂtransferase activity. Next, because CTB is known to trigger
the pathway involved in regulating <i>Sox9</i>, a master
regulator of chondrogenic differentiation, we modifed a <i>Sox9</i>-specific NanoScript with CTB to enhance chondrogenic gene activity
and differentiation. Because NanoScript is a tunable and robust platform,
it has potential for various gene-regulating applications, such as
stem cell differentiation