Stemgent® mRNA Reprogramming Factors Set: hOKSML (1 Set)_详细资料

产品信息

供应商： Stemgent 英文名： Stemgent® mRNA Reprogramming Factors Set: hOKSML (1 Set) 保质期： 1年保存条件： -80℃ 规格： 1 Set

Stemgent致力于通过提供由一些世界DJ干细胞科研人员所开发的专有试剂盒工具促进干细胞科学的发展，Stemgent的产品包括：针对多能性、自我更新和分化的小分子；mRNA转录因子；基质；细胞系；细胞因子；抗体等。

Product Overview
Stemgent mRNA Reprogramming Factors Set: hOKSML contains mRNA encoding Oct4, Klf4, Sox2, c-Myc, Lin-28, and nuclear GFP (nGFP). These in vitro transcribed mRNAs incorporate both pseudouridine and 5-methylcytidine-modified nucleotides to minimize the cellular interferon response to single stranded RNA.^1,2Repeated transfection of mRNA encoding Oct4, Klf4, Sox2, c-Myc and Lin-28 has been shown to reprogram human fibroblasts.² nGFP mRNA is included for use as a positive control for RNA transfection during the cellular reprogramming process.
Advantages

Faster reprogramming, colonies emerge as early as 16 days

Stoichiometric control of individual reprogramming factors

Eliminates need for backend screening of viral systems

Validated protocol using normal and diseased cells
Product Specifications
Kit / Set Components
2 vials - Stemgent Oct4 mRNA, Human (Cat. No. 05-0014), 20 μg
1 vial - Stemgent Klf4 mRNA, Human (Cat. No. 05-0015), 20 μg
1 vial - Stemgent Sox2, Human (Cat. No. 05-0016), 20 μg
1 vial - Stemgent c-Myc mRNA, Human (Cat. No. 05-0018), 20 μg
1 vial - Stemgent Lin-28 mRNA, Human (Cat. No. 05-0017), 20 μg
1 vial - Stemgent nGFP mRNA (Cat. No. 05-0019), 20 μg
Formulation
100 ng/μl in RNase-free TE buffer, pH 7.0
Storage and Stability
Store at or below -70°C. Stable for a minimum of 3 months from date of receipt when stored as directed. Wear gloves when handling this product. Use RNase-free tubes and barrier pipette tips. Thaw on ice. Keep on ice until the sample is removed. Avoid multiple freezing and thawing cycles (<5). Aliquot to experimental quantities.
Quality Control
Proper protein expression and localization have been confirmed by immunocytochemistry for reprogramming factors and by fluorescence microscopy for nGFP.
Recommended Usage
mRNAs should be transfected using a protocol specifically optimized for the cell type used in the experiment.
Related Products

Stemgent^® Oct4 mRNA, human (Cat.No.05-0014 )

Stemgent^® Klf4 mRNA, Human (Cat.No.05-0015)

Stemgent^® Sox2 mRNA, Human (Cat.No.05-0016)

Stemgent^® c-Myc mRNA, Human (Cat.No.05-0018)

Stemgent^® Lin-28 mRNA, Human (Cat.No.05-0017)

Stemgent^® nGFP mRNA (Cat.No.05-0019)

Pluriton™ mRNA Reprogramming Medium (Cat.No. 00-0070)

Stemfect™ RNA Transfection Kit (Cat.No.00-0069)

Stemolecule™ Y27632 (Cat.No.04-0012)

Stemfactor™ Fibroblast Growth Factor-basic (Human Recombinant) (Cat.No.03-0002)
产品特点
1.快速 Fast
Stemgent® MicroRNA-Enhanced mRNA Reprogramming System只需2周即可将靶细胞重编程为iPS细胞，与仙台病毒（Sendai Virus）、非整合型DNA载体（Episomal DNA Vectors）等传统iPS重编程方法相比，缩短实验周期超过50%以上（Figure. 1）。
Figure 1. Stemgent® mRNA重编程系统、仙台病毒依赖的重编程系统以及游离型DNA载体依赖的重编程系统重编程时程比较。

简便 Easy
与病毒、DNA载体依赖的iPS重编程手段不同，使用Stemgent® MicroRNA-Enhanced mRNA Reprogramming System对靶细胞进行重编程，生成的iPS细胞无需进行复杂耗时的下游筛选程序，只需简简单单6个步骤，即可轻松构建iPS细胞系。

Description Day

Step 1 Material Preparation Prior to starting

Step 2 Plate Cells 0

Step 3 Transfect Cells 1 to 12

Step 4 Identify Cells 13-14

Step 5 Pick and Passage iPS Cell Colonies 15-16

Step 6 Maintain iPS Cell Cultures 16+

注：因所用细胞类型及实验条件不同，上述时间轴可能略有不同，仅供参考。

Figure 2. Stemgent® MicroRNA-Enhanced mRNA Reprogramming System重编程过程时间表、显微镜下观察到的重编程过程中靶细胞形态变化。第12天，形成的iPS克隆表达多能型Marker TRA-1-81。

GX Efficient
病毒、DNA载体等依赖的iPS重编程技术，其重编程效率介于0.00001%到1%之间不等，而Stemgent® MicroRNA-Enhanced mRNA Reprogramming System的重编程效率远远大于1%，极大地提高了iPS克隆的产出率。
表1. 不同类型iPS重编程手段效率比较

Reprogramming Method Efficiency Integrating Screening

RNA >1% No No

仙台病毒（Sendai virus） 0.01-1% No Yes

DNA载体（Episomal/Minicircle） 0.00% Possible Yes

慢病毒（Lentivirus） 0.001-0.01% Yes Yes

腺病毒（Adenovirus） 0.0001-0.001% Possible Yes

Protein 0.00% No No

某些iPS研究领域，例如iPS临床研究、ZL应用等，对iPS细胞的安全性提出了极高的要求。使用Stemgent® MicroRNA-Enhanced mRNA Reprogramming System重编程靶细胞，无需担心构建的iPS细胞系可能含有病毒载体残留，亦无需担心无关的基因整合进宿主基因组，是目前适用于临床研究及ZL的最理想选择。安全 Safe
而以仙台病毒（Sendai Virus）为载体的iPS重编程手段，其病毒基因组虽然不会整合进宿主染色体，仙台病毒本身也不具备复制能力，但是为了构建真正意义上的virus-free的iPS细胞系，通常需要经过一个10-20代、耗时费力的筛选过程，以确保ZZ得到的iPS细胞系无病毒载体残留。以慢病毒（Lentivirus）体的iPS重编程手段，病毒基因组能够整合进宿主基因组，会给后续的iPS细胞分析带来干扰。

高性价比 Cost Effective
以仙台病毒为载体的iPS重编程试剂盒非常昂贵，综合重编程效率、时间及劳动力成本、安全性等多重考虑，Stemgent® MicroRNA-Enhanced mRNA Reprogramming System无疑是性价比的上乘之选。

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	Description	Day
Step 1	Material Preparation	Prior to starting
Step 2	Plate Cells	0
Step 3	Transfect Cells	1 to 12
Step 4	Identify Cells	13-14
Step 5	Pick and Passage iPS Cell Colonies	15-16
Step 6	Maintain iPS Cell Cultures	16+

Reprogramming Method	Efficiency	Integrating	Screening
RNA	>1%	No	No
仙台病毒（Sendai virus）	0.01-1%	No	Yes
DNA载体（Episomal/Minicircle）	0.00%	Possible	Yes
慢病毒（Lentivirus）	0.001-0.01%	Yes	Yes
腺病毒（Adenovirus）	0.0001-0.001%	Possible	Yes
Protein	0.00%	No	No