质量水平
产品线
MISSION®
浓度
≥1x106 VP/ml (via p24 assay)
技术
capture ELISA: 106 TU/mL using p24
运输
dry ice
储存温度
−70°C
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一般描述
想要查看更多应用数据、实验方案和载体图谱,请访问 sigma.com/shrna。
Small interfering RNAs (siRNAs) expressed from short hairpin RNAs (shRNAs) are a powerful way to mediate gene specific RNA interference (RNAi) in mammalian cells. The MISSION product line is based on a viral vector-based RNAi library against annotated mouse and human genes. shRNAs that generate siRNAs intracellularly are expressed from amphotropic lentivirus viral particles, allowing screening in a wide range of mammalian cell lines. In these cell lines, MISSION shRNA clones permit rapid, cost efficient loss-of-function and genetic interaction screens.
The lentiviral transduction particles are produced from an shRNA lentiviral non-target control plasmid. It is useful as a negative control in experiments with the MISSION shRNA target sets.
Unlike murine-based MMLV or MSCV retroviral systems, lentiviral-based particles permit efficient infection and integration of the specific shRNA construct into differentiated and non-dividing cells, such as neurons and dendritic cells,1 overcoming low transfection and integration difficulties when using these cell lines. Self-inactivating replication incompetent viral particles are produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids.2-3
In addition, the lentiviral transduction particles are pseudotyped with an envelope G glycoprotein from vesicular stomatitis virus (VSV-G), allowing transduction of a wide variety of mammalian cells.4 The lentiviral transduction particles are titered via a p24 antigen ELISA assay and pg/ml of p24 are then converted to transducing units per ml using a conversion factor. The conversion can be viewed at: www.tronolab.com.
应用
法律信息
储存分类代码
12 - Non Combustible Liquids
WGK
WGK 3
闪点(°F)
Not applicable
闪点(°C)
Not applicable
个人防护装备
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
法规信息
商品
Methods for lentiviral transduction of Jurkat cells were compared. Spinoculation was compared with overnight incubation with polybrene (hexadimethrine bromide) and fibronection-coated plates.
实验方案
Detailed procedure for how to perform a lentiviral transduction of MISSION shRNA lentiviral particles to achieve a stable long term silencing and phenotypic change.
This detailed procedure allows you to transduce Mouse Embryonic Fibroblasts (MEF) using MISSION ExpressMag Super Magnetic Kit.
如何利用MISSION shRNA慢病毒颗粒进行慢病毒转导,以实现稳定的长期沉默和表型变化的详细操作步骤。
相关内容
Instructions
This protocol describes the use of MISSION TRCshRNA Lentiviral Particles and provides a system for long-term silencing andphenotypic observation.
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