一般描述
EMD Millipore’s Simplicon RNA Reprogramming Kit is a safe and efficient method to generate integration free, virus-free human iPS cell using a single transfection step. The technology is based upon a positive strand, single-stranded RNA species derived from non-infectious (non-packaging), self-replicating Venezuelian equine encephalitis (VEE) virus3. The Simplicon RNA replicon is a synthetic in vitro transcribed RNA expressing all four reprogramming factors (OKS-iG; Oct4, Klf4, Sox2 and Glis1) in a polycistronic transcript that is able to self-replicate for a limited number of cell divisions.
应用
• Integration-free, footprint-free iPS cells. No risk of genomic integration
• Safe, virus free, synthetic polycistronic RNA replicon (all four reprogramming factors in 1 RNA strand)
•Only 1-day transfection required. The RNA replicon is able to self-replicate, elliminating the need for laborious daily transfection of multiple individual mRNAs over a 14 day period.
• Efficient and rapid reprogramming.
• No screening required to ensure viral remnants are not present.
• Controlled elimination of synthetic RNA replicon by removal of B18R protein.
• Validated for reprogramming in feeder-based and feeder-free culture conditions.
Stem Cell Research
制备说明
• Human recombinant B18R protein: Stable for 4 months from date of receipt when stored appropriately at -80°C. For best recovery, quick-spin the vial prior to opening. Thaw on ice. While on ice, aliquot B18R protein into sterile, nuclease-free, low protein-binding eppendorf tubes and store at -80°C. B18R protein must be kept on ice in order to avoid degradation. Limit repeated freeze-thaw cycles. Use in a sterile RNAse-free environment.
• Human iPS Reprogramming Boost Supplement II: Stable for 4 months at -20˚C from date of receipt. Upon first thaw, aliquot into smaller working volumes and freeze at -20°C. Upon addition of the small molecule components to the media, filter the supplemented media with a 0.22 μM filtration unit and stored at 2-8˚C. For optimal results, prepare sufficient supplemented media for a 1 week supply of media changes.
分析说明
其他说明
2. B18R RNA: (Part No. CS210584) One (1) vial containing 10 µL of RNA (1 ug/uL). Store at -80°C.
3. Human Recombinant B18R Protein, Carrier-Free: (Part No. GF156) One (1) vial containing 50 ug of 0.5 mg/mL stock of B18R protein. Store at -80°C.
4. Human iPS Reprogramming Boost Supplement II: (Part No. SCM094). Store at -20°C.
法律信息
免责声明
储存分类代码
10-13 - German Storage Class 10 to 13
WGK
WGK 2
商品
Human iPSC neural differentiation media and protocols used to generate neural stem cells, neurons and glial cell types.
The Simplicon™ RNA Reprogramming Technology is a next generation reprogramming system that uses a single synthetic, polycistronic self-replicating RNA strand engineered to mimic cellular RNA to generate human iPS cells.
Fibroblast growth factors (FGFs) regulate developmental pathways and mesoderm/ectoderm patterning in early embryonic development.
实验方案
Stem cell reprogramming protocols to generate human induced pluripotent stem cells (iPSCs) including viral and non-viral RNA based methods.
产生人诱导多能干细胞（iPSC）的干细胞重编程实验方案，包括基于病毒和非病毒RNA的方法。
相关内容
"Aging: getting older, exhibiting the signs of age, the decline in the physical (and mental) well-being over time, leading to death. Since the beginning of time, man has been obsessed with trying to slow down, stop, or even reverse the signs of aging. Many have gone as far as experimenting with nutritional regimens, eccentric exercises, fantastic rituals, and naturally occurring or synthetic wonder-elements to evade the signs of normal aging. Biologically speaking, what is aging? And what does the latest research tell us about the possibility of discovering the elusive “fountain of youth”? Many advances in our understanding of aging have come from systematic scientific research, and perhaps it holds the key to immortality. Scientifically, aging can be defined as a systems-wide decline in organismal function that occurs over time. This decline occurs as a result of numerous events in the organism, and these events can be classified into nine “hallmarks” of aging, as proposed by López-Otin et al. (2013). Several of the pathologies associated with aging are a direct result of these events going to extremes and may also involve aberrant activation of proliferation signals or hyperactivity. The hallmarks of aging have been defined based on their fulfillment of specific aging related criteria, such as manifestation during normal aging, acceleration of aging if experimentally induced or aggravated, and retardation of aging if prevented or blocked, resulting in increased lifespan. The nine hallmarks of aging are genomic instability, telomere attrition, epigenetic alterations, loss of proteostasis, deregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication. The biological processes underlying aging are complex. By understanding the hallmarks in greater detail, we can get closer to developing intervention strategies that can make the aging process less of a decline, and more of a recline."
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