Allele Biotech-Introducing Cost Effectiveness to Research
I
n 2010, Warren et al. illustrated a footprint free method to
reprogram adult cell to iPSCs using modied mRNA mol
-ecules. This method was groundbreaking for two reasons,
the use of modied RNA bases mitigated cell immune re
-
sponse to transfected mRNA, and the efciency 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
T
he Stealth Express® Neural Transcription Factors sup
-
port footprint-free induction using mRNA transfection, a new reprogramming technology that’s faster and more ef
-
cient than conventional viral-based approaches. Products
available include in vitro transcription templates for large-
scale 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 anked by 5’ and 3’ untranslated regions de
-
signed to maximize the translational efciency and cytoplas
-
mic stability of the mRNA and validated in the iPSC applica
-
tion. The templates drive incorporation of 120-nt polyA tails into RNA transcripts, eliminating the need to enzymatically polyadenylate IVT products before use. Also offered seper
-
ately are: modied UTP and CTP, standard ATP and GTP, T7 polymerase, ARCA, DNase, and RNA Purication Kit. The Stealth Express®Neural 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. ♦ Templates contain the T7 phage polymerase promoter al
-
lowing 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 syn
-
thetic mRNA
Neural Transcription Factors
, exogenous
expression of Myt 1l, Ascl1, and Neuro D2 via mRNA transfection for directing cell fate into neuronal cells.
Description
Box 1 | Product List
iPS Induction System Content
DescriptionCat. No. Qty
Neural Transcription Factors PCR Template Set (Myt1l, Ascl1, Neuro D2 and mWasabi) ABP-SC-SENTFSET
10 µg x 4
Myt1l
IVT Template
ABP-FP-SEMYT1
10 µg
Ascl1
IVT Template
ABP-FP-SEASC1
10 µg
Neuro D2
IVT Template
ABP-FP-SENED2
10 µg
mWasabi IVT Template
ABP-FP-SEMWASABI
10 µg
2.5X Nucleotide Mix w/ Cap AnalogABP-PP-NTPMIX
100 µl
Surface Bind RNA Purication KitABP-PP-SBRNA02525 RXNS
OR
RNA Purication SystemABP-PP-RNAPUR02525 RXNSLanYFP IVT Template
Coming Soon10 µg
Functional Proteins
Cre IVT Template
ABP-RP-SECRE
10 µg
Luciferase IVT TemplateABP-RP-SELUC
10 µg
Features
F
or Research Use Only. Not for Diagnostic or Therapeutic Use.
Purchase does not include or carry any right to re
-sell 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
Allele Biotech-Introducing Cost Effectiveness to Research
I
n order to efciently 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 modications 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 repro-gramming.
Box 2 | Stealth Express Overview
ComponentCat. No.Description
2.5x Nucleotide Mixw/ Cap Analog
ABP-PP-NTPMIX
Blend of components needed
for IVT reactions. Included in the
Ready-Mix cocktail:
Modifed UTP (Pseudouridine)Modifed CTP (5-Methylcytidine) ATP GTP ARCA (Anti-Reverse Cap Analog)
Modied bases are intended to
ameliorate immunological respons-
es towards the mRNA product.
SurfaceBind RNA Purication
Kit
ABP-PP-SBRNA025
For purication of RNA products
Transfection Reagent
Coming Soon
Cationic vehicle to facilitate endo-
cytic delivery of mRNA.
B18
Coming Soon
Immune inhibitor allows for repeat
mRNA transfections.
Stem Cell Media
Coming Soon
Cell media optimized for the cultur-ing of ES and iPS cells.
Overview
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