Introduction CRISPR / system CAS9, which can be utilized in vitro biological experiments have recently caught a lot of attention for their important role and benefits. However, realization of the full potential of CRISPR / CAS9 approach requires addressing the various challenges and side effects. Expression gene and the potential side effects of the CRISPR / CAS9 in Human cells that are still to be explained.
The aim of our study was to explore the effect of CRISPR / CAS9 on gene expression in 293T cells. A Cas9-expressing plasmid PX458 and PX458 Cas9-T2A-off plasmids used to study the role of CRISPR / CAS9 at set gene expression in 293T cells. Gene expression in 293T cells after transfection of plasmid PX458 or PX458-T2A plasmid RNA was detected by sequencing and correlative statistical analysis. Differential gene expression in transfected 293T second PX458 and PX458-T2A cells transfected 293T 293T cells compared with normal cells detected using chain quantitative reverse transcription polymerase reaction (RT qPCR). MRNA and protein levels were measured using reverse transcription PCR and Western blot.
Co-IP assay combined with a shotgun LC-MS / MS is used to investigate differences NGFR protein-interaction between PX458 and PX458 transfected 293T cells transfected 293T cell-T2A. In this study, we observed that the PX458 plasmid transfection and expression Cas9 can affect the expression of different gene , including FOSB (FBJ murine osteosarcoma virus Onco gene homolog B), IL – 11 (Interleukin- 11 ), MMP1 (matrix metalloproteinases), CYP2D6 (CytochromeP4502D6), and NGFR (matrix metalloproteinase 1).
Cas9 Protein Triggers Differential Expression of Inherent Genes Especially NGFR Expression in 293T Cells.
Downregulation of NGFR after transfection PX458 confirmed by RT qPCR and western blot analysis. NGFR expression was significantly lowered in PX458 transfected 293T 293T cells than in normal cells and PX458-T2A transfected 293T cells. Dilution co-IP was analyzed with a shotgun LC-MS / MS showed total 183 proteins interact with NGFR in 293T cells transfected PX458 while 221 proteins interact with NGFR-T2A PX458 identified in transfected 293T cells using machine mascot.
Cas9 expression by transfection of plasmid PX458 negatively correlated with the level NGFR NGFR mRNA and protein expression in 293T cells, while the PX458-T2A, where Cas9 disabled, does not affect the expression of NGFR. Decreased expression of NGFR also affects the number of proteins that interact with NGFR. These results indicate that the effect Cas9 in NGFR expression and other expressions gene should be considered when developing cell-based research and therapy utilizing the CRISPR / CAS9 system.
Profile inflammation and osteogenic hematoma Fracture Human Potential. form of fracture hematoma immediately after the fracture and is considered important for bone healing process. molecular composition has been briefly investigated the existence of our current understanding is based on animal studies.
Description: A sandwich quantitative ELISA assay kit for detection of Canine Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Canine Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Chicken Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Chicken Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Equine Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Equine Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Human Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Human Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Mouse Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Mouse Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Porcine Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Porcine Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Rat Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Rat Interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: Quantitativesandwich ELISA kit for measuring Dog Interleukin-13 (IL13) in samples from serum, plasma, tissue homogenates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.
Description: Quantitativesandwich ELISA kit for measuring Dog Interleukin-13(IL13) in samples from serum, plasma, tissue homogenates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: Quantitativesandwich ELISA kit for measuring Sheep interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.
Description: Quantitativesandwich ELISA kit for measuring Sheep interleukin 13 (IL13) in samples from serum, plasma, tissue homogenates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: Human IL-13 ELISA development kit contains the key components required for the quantitative measurement of natural and/or recombinant IL-13 in a sandwich ELISA format. Using the ELISA protocol described below, this kit provide ssufficient reagents to assay IL-13 in approximately 1,500 ELISA plate wells.
The purpose of this study to analyze the content of inflammatory cytokines in the fracture hematoma Human and determine their effect on osteoprogenitor cells. Twenty-three patients were recruited following informed consent. peripheral blood, and bone fracture hematoma collected. A Luminex assay at the level of 34 cytokines done and autologous peripheral blood samples served as controls. Mesenchymal Stem Cells (MSCs) were isolated following collagenase digestion and functional tests performed. Gene expression analysis of 84 key osteogenic molecules do.
