NanoScript: A Nanoparticle-Based Artificial Transcription Factor for Effective Gene Regulation

Abstract

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.

Peptide-functionalized gold nanoparticles: versatile biomaterials for diagnostic and therapeutic applications

“…Lee et al conjugated separately to GNPs peptide sequences representing the three essential domains of natural TFs, namely a nuclear localisation signal, a DNA binding domain and an activation domain 90 . The resulting construct, named NanoScript, was co-transfected with an alkaline phosphatase (ALP) reporter plasmid into HeLa cells.…”

Towards combinatorial transcriptional engineering

“…Recently, Blackburn et al, (2016) could synthesize and characterize 400 ZNFs in a short duration by integrating gene synthesis and microfluidic analysis platforms. When augmented by efficient delivery systems like the nanoparticle based technology devised by Patel et al, (2014), similar technologies will expand the application of ZNFs and TALEs in synthetic biology. An easier alternative for controlling multiple nodes of a complex genetic circuit is the CRISPR/Cas9 system due to the relative simplicity in designing and targeting gRNAs to different genomic targets.…”

New Approaches to Colorectal Cancer Treatment; An Overview

“…There are monoclonal antibodies such as specific anti-human CD133 mAb, anti human CD29 monoclonal antibody-clone 12G10, SWA11 mAb and KM4056 mAb for CD133, CD29 , CD24 and LGR5 antigens respectively. For a more effective and specific drug delivery, it is suggested to target colorectal cancer stem cells with specific antibody conjugated with nanoparticles which contain loaded drug (15).…”

“…Nano Script has two interesting features that make it appropriate platform in the field of stem cells: First, it is non-viral, that is a good alternative to viral vectors. Second, simple arrangement of the sequence of any molecule on Nano Script is possible so they can specifically target and induce differentiation (15). Liu et al developed a new nanostructure which contains a combination of cross-linked hydrophilic polymers with lipids.…”