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Cas9 Protein Triggers Differential Expression of Inherent Genes Especially NGFR Expression in 293T Cells.

Cas9 Protein Triggers Differential Expression of Inherent Genes Especially NGFR Expression in 293T Cells.
Posted by Rachael

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.
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.

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Recombinant Rhesus Macaque Interleukin-13 protein(IL13) (Active)

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Recombinant Mouse Interleukin-13/IL-13 Protein (Active)

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Description: Mouse

Recombinant Mouse Interleukin-13/IL-13 Protein (Active)

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Active Interleukin 10 (IL10)

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Description: Recombinant Human Interleukin 10 expressed in: 293F cell

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Description: Recombinant Human Interleukin 11 expressed in: E.coli

Active Interleukin 11 (IL11)

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Active Interleukin 11 (IL11)

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Description: Recombinant Human Interleukin 15 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

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Active Interleukin 17 (IL17)

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Description: Recombinant Human Interleukin 18 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

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Description: Recombinant Mouse Interleukin 18 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

Active Interleukin 19 (IL19)

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Active Interleukin 19 (IL19)

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Description: Recombinant Human Interleukin 21 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

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Active Interleukin 21 (IL21)

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Description: Recombinant Human Interleukin 25 expressed in: E.coli

Active Interleukin 25 (IL25)

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EUR 280

Active Interleukin 25 (IL25)

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Active Interleukin 33 (IL33)

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Description: Recombinant Human Interleukin 33 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

Active Interleukin 34 (IL34)

APC007Hu01 10ug
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Description: Recombinant Human Active Interleukin 34 (IL34) expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

Active Interleukin 34 (IL34)

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EUR 316

Active Interleukin 34 (IL34)

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Active Interleukin 34 (IL34)

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Active Interleukin 35 (IL35)

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EUR 200

Active Interleukin 35 (IL35)

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Description: Recombinant Human Interleukin 35 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

Active Interleukin 29 (IL29)

APC029Hu01 10ug
EUR 211

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4-APC029Hu01
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Description: Recombinant Human Interleukin 29 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

Active Interleukin 29 (IL29)

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Active Interleukin 22 (IL22)

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Description: Recombinant Human Interleukin 22 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

Active Interleukin 22 (IL22)

APC032Hu61 10ug
EUR 324

Active Interleukin 22 (IL22)

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Active Interleukin 20 (IL20)

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EUR 219

Active Interleukin 20 (IL20)

4-APC058Hu01
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Description: Recombinant Human Interleukin 20 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

Active Interleukin 20 (IL20)

APC058Mu61 10ug
EUR 371

Active Interleukin 24 (IL24)

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EUR 211

Active Interleukin 24 (IL24)

4-APC064Hu01
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Description: Recombinant Human Interleukin 24 expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

Active Interleukin 23 (IL23)

APX262Mu61 10ug
EUR 340

Active Interleukin 18 (IL18)

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EUR 739.2

Active Interleukin 18 (IL18)

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Active Interleukin 18 (IL18)

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Active Interleukin 21 (IL21)

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EUR 997.7

Active Interleukin 21 (IL21)

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EUR 3893.5

Active Interleukin 21 (IL21)

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EUR 599.1

Active Interleukin 33 (IL33)

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EUR 1013.1

Active Interleukin 33 (IL33)

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EUR 5116.8

Active Interleukin 33 (IL33)

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EUR 787.2

Active Interleukin 34 (IL34)

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EUR 997.7

Active Interleukin 34 (IL34)

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EUR 3931.2

Active Interleukin 34 (IL34)

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EUR 604.8

Active Interleukin 29 (IL29)

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EUR 651.2

Active Interleukin 29 (IL29)

RPU55746-1mg 1mg
EUR 3294.2

Active Interleukin 29 (IL29)

RPU55746-50ug 50ug
EUR 506.9

Active Interleukin 22 (IL22)

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EUR 1282.6

Active Interleukin 22 (IL22)

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EUR 5054.4

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.

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