Neural Transcription Factors,

exogenous expression of Myt 1l, Ascl1, and Neuro D2 via mRNA transfection for directing cell fate into neuronal cells.

n 2010, Warren et al. illustrated a footprint free method to reprogram adult cell to iPSCs using modified mRNA molecules. This method was groundbreaking for two reasons, the use of modified RNA bases mitigated cell immune response to transfected mRNA, and the efficiency of induction was higher than viral transfection. The next progression of these studies are begining to focus on directing the cell fate of iPSC into the desired cell types. Myt1l, Ascl1 and Neuro D2 are transcription factors found to reprogram stem cells into functional induced neuronal (iN) cells

Box 1 | Product List

iPS Induction System Content
Description Neural Transcription Factors PCR Template Set (Myt1l, Ascl1, Neuro D2 and mWasabi) Myt1l IVT Template Ascl1 IVT Template Neuro D2 IVT Template mWasabi IVT Template 2.5X Nucleotide Mix w/ Cap Analog Surface Bind RNA Purification Kit RNA Purification System LanYFP IVT Template Cat. No. ABP-SC-SENTFSET Qty
10 g x 4

ABP-FP-SEMYT1 ABP-FP-SEASC1 ABP-FP-SENED2 ABP-FP-SEMWASABI ABP-PP-NTPMIX ABP-PP-SBRNA025

OR

10 g 10 g 10 g 10 g 100 l

Description
he Stealth Express Neural Transcription Factors support footprint-free induction using mRNA transfection, a new reprogramming technology thats faster and more efficient than conventional viral-based approaches. Products available include in vitro transcription templates for largescale synthesis of mRNA encoding three neural factors associated with neuronal cell fate induction (Myt1l, Ascl1, Neuro D2). Linear DNA templates are ready to use directly in IVT reactions. Within each template, the coding sequence for a protein is flanked by 5 and 3 untranslated regions designed to maximize the translational efficiency and cytoplasmic stability of the mRNA and validated in the iPSC application. The templates drive incorporation of 120-nt polyA tails into RNA transcripts, eliminating the need to enzymatically polyadenylate IVT products before use. Also offered seperately are: modified UTP and CTP, standard ATP and GTP, T7 polymerase, ARCA, DNase, and RNA Purification Kit. The Stealth ExpressNeural Transcription Factors gives the ability to create and purify mRNA transcripts encoding the 3 neural transcription factors, and then transfect them into target cells for reprogramming into iNC.

25 RXNS 25 RXNS 10 g

ABP-PP-RNAPUR025 Coming Soon

Functional Proteins
Cre IVT Template Luciferase IVT Template ABP-RP-SECRE ABP-RP-SELUC 10 g 10 g

Features
Templates contain the T7 phage polymerase promoter allowing the widely available T7 polymerase to be used. Inherently non-mutagenic IVT template for footprint-free gene expression Eliminates safety concerns and bio-containment issues associated with virus Potential to induce iPSC into functional induced neuronal (iN) cells Template can be used to make over two milligrams of synthetic mRNA

or Research Use Only. Not for Diagnostic or Therapeutic Use. Purchase does not include or carry any right to resell or transfer this product either as a stand-alone product or as a component of another product. Any use of this product other than the permitted use without the express written authorization of Allele Biotech is strictly prohibited

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Overview

n order to efficiently and effectively reprogram cells into iPSC using mRNA as a method of gene delivery, multiple steps must be taken to ensure that:

mRNA is successfully invitro-transcribed with proper modifications in effort to suppress target cell immune activity towards mRNA molecules. mRNA is of suitable purity mRNA products are delivered into target cells Allele Biotech conveniently offers imperative components to produce footprint free iPSC as described in Warren et al.2010 publication. The following table outlines necessary components required, in addition to the iPS IVT templates, for cell reprogramming.

Box 2 | Stealth Express Overview

Component 2.5x Nucleotide Mix w/ Cap Analog

Cat. No. ABP-PP-NTPMIX

Description Blend of components needed for IVT reactions. Included in the Ready-Mix cocktail:
Modified UTP (Pseudouridine) Modified CTP (5-Methylcytidine) ATP GTP ARCA (Anti-Reverse Cap Analog)

Modified bases are intended to ameliorate immunological responses towards the mRNA product.

SurfaceBind RNA Purification Kit Transfection Reagent B18 Stem Cell Media

ABP-PP-SBRNA025 Coming Soon Coming Soon Coming Soon

For purification of RNA products Cationic vehicle to facilitate endocytic delivery of mRNA. Immune inhibitor allows for repeat mRNA transfections. Cell media optimized for the culturing of ES and iPS cells.

Allele Biotech-Introducing Cost Effectiveness to Research