货号: 05-0029
供应商: Stemgent
英文名: Stemgent® MyoD mRNA, Human (20 µg)
保质期: 1年
保存条件: -80℃
规格: 20 µg

Stemgent致力于通过提供由一些世界顶尖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

The Stemgent MyoD mRNA encodes the human myogenic differentiation (MYOD) protein. This in vitro transcribed mRNA incorporates both pseudouridine and 5-methylcytidine modified nucleotides to minimize the cellular interferon response to single stranded RNA1,2. MyoD is a member of the basic helix-loop-helix (bHLH) family of muscle-specific transcription factors. Over expression of MyoD has been shown to convert fibroblasts and numerous other cell types directly into skeletal muscle3,4.

Product Specifications

Size

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 6 months from date of receipt, when stored as directed. Wear gloves when handling this product. Use RNase-free reagents, tubes and barrier pipette tips. Thaw and keep on ice until the sample is removed. Avoid multiple freezing and thawing cycles. Aliquot to experimental quantities.

Quality Control

Concentration, RNA purity, molecular weight, sterility, as well as proper protein expression and localization have been confirmed.

Recommended Usage

Stemgent MyoD mRNA should be transfected with Stemfect™ RNA Transfection Reagent and optimized according to the target cell type used in the experiment. Please reference Protocol: MyoD mRNA, Human.

Notice To Purchaser

 

Purchaser represents and warrants that it will use MyoD mRNA, Human purely for research purposes, not for diagnostic use, not for resale, and not for use in humans or veterinary applications. Stemgent will not be held responsible for patent infringement or other violations that may occur with the use of our products.

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)。

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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。

 DescriptionDay
Step 1Material PreparationPrior to starting
Step 2Plate Cells0
Step 3Transfect Cells1 to 12
Step 4Identify Cells13-14
Step 5Pick and Passage iPS Cell Colonies15-16
Step 6Maintain iPS Cell Cultures16+
 

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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。

 

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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。

 

高效 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 MethodEfficiencyIntegratingScreening
RNA>1%NoNo
仙台病毒(Sendai virus0.01-1%NoYes
DNA载体( Episomal/Minicircle 0.00%PossibleYes
慢病毒(Lentivirus0.001-0.01%YesYes
腺病毒(Adenovirus0.0001-0.001%PossibleYes
Protein0.00%NoNo

某些iPS研究领域,例如iPS临床研究、治疗应用等,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治疗的最理想选择。 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,以确保最终得到的iPS细胞系无病毒载体残留。以慢病毒(Lentivirus)体的iPS重编程手段,病毒基因组能够永久整合进宿主基因组,会给后续的iPS细胞分析带来干扰。

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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。 

温馨提示:不可用于临床治疗。