Development of a High Efficiency Transfection Reagent for CRISPR/Cas9-Genome Editing In Human Induced Pluripotent Stem Cells and iPSC-derived Neural Stem Cells - NYSCF 2016
Here, human iPSCs and NSCs were transfected as adherent cells or in suspension with a new flexible CRISPR/Cas9 gene delivery reagent to overcome inhibition of transfection observed in the center of colonies grown in traditional culture systems. Download full poster to learn more.
Development of an Enabling System for High Efficiency Reprogramming and CRISPR-Based Genome Editing - ISSCR 2016
In order to streamline the workflow and increase throughput while simultaneously enhancing accuracy, we have developed an integrated transfection and culture system in which cells can be reprogrammed and the resultant iPSC can undergo genome editing.
Cas9 mRNA Delivery: An Approach for Efficient Transfection with Modified mRNA CRISPR - Congress Boston '16
In this study, a new mRNA-In® CRISPR Transfection Reagent was used to transfect Cas9 mRNA into a variety of cell types. Transfection with mRNA-In® CRISPR resulted in >95% transfection efficiency in studied cells.
Titration of mRNA Delivery: An Approach for Efficient Transfection with Modified or Unmodified mRNA
In this study, four different cell types (HeLa, human neural stem, HUVEC and adipose-derived mesenchymal cells) were transfected with various amounts of either modified 5meC, Ψ GFP mRNA or unmodified mRNA.The data indicates that mRNA transfection is possible for both modified and unmodified mRNA, with >95% transfection efficiency and minimal toxicity at amounts of up to 500 ng mRNA across all four cell types.
Nucleotide Modifications for Improved Messenger RNA Expression
In the last several years, in vitro transcribed mRNA has gained traction as a potential new therapeutic agent to deliver genetic information.mRNA provides several inherent benefits over traditional plasmid- and virus-based methods for gene therapy.
Making the Optimal Messenger RNA for Gene Therapy Applications
Recently, there has been significant interest in the use of messenger RNA (mRNA) based expression systems for gene therapy applications. Several groups have shown that mRNA is an attractive vehicle for therapeutic gene expression in mammals (Kormann et al. Nat. Biotechnol (2011) 29, 154; Kariko et al. Molecular Therapy (2012) 20, 948). mRNAs are expressed in the cytoplasm of cells which may improve expression in non-dividing cells, which are difficult to transfect.
A Reproducible, Efficient and User-friendly RNA Reprogramming System for Footprint-free Human iPSC Generation
Induced pluripotent stem cells (iPSCs) are becoming an invaluable tool for disease modeling and drug discovery, but the prospect of applying iPSCs in cell therapy is still greatly limited by the risk of genomic alterations using viral methods for gene delivery during the reprogramming process. Recent advances...
Functional Synaptic Activity in Human Stem Cell Derived Neurons
Neurons derived from human pluripotent stem cells (hPSCs) hold great potential as a physiologically relevant platform for studying human responses to neuromodulatory agents. However, methods to reproducibly generate synaptically active populations with emergent network behaviors have not been described...
Human Induced Pluripotent Stem Cell-Derived Neurons as a Reproducible System for Drug Discovery
Neurotoxicity studies and compound screening for drug candidates for neurological and neurodegenerative disorders rely on mature cultures of neurons and other neural cells. Currently, primary rodent neurons are the widely accepted norm in drug screening and toxicity assays.
Development of High Throughput Synaptic Functional Assays in Neurons Derived from Human Induced Pluripotent Stem Cells and Characterization of Pharmacological Responses
Use of human neurons for neurotransmission screening applications requires that cultures achieve a sufficient degree of synaptic maturation to yield a measurable proportion of synapses with pre- and post-synaptic functionality. Here, we show that cultures of neurons derived from human iPSCs can be utilized in the MANTRA system for high-throughput assays to measure evoked Ca+2 transients or evoked pre-synaptic responses.
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