Plasmid Preparation:Article Title: CRB2 depletion induces YAP signaling and disrupts mechanosensing in podocytes.
Article Snippet: .. 160 Briefly, 15 μg of vector plasmid, 10 μg of psPAX2 packaging plasmid (Addgene #12260 161 generated in Dr Didier Trono’s laboratory, EPFL, Switzerland), 5 μg of pMD2.G envelope 162 plasmid (Addgene #12259, generated in Dr Trono’s laboratory) and 2.5 μg of pRSV-Rev 163 plasmid (Addgene #12253, generated in Dr Trono’s laboratory) were transfected into 293 T 164 cells. .. Vector particles were collected from the filtered conditioned medium at 72 h post-165 transfection.
Article Title: Myotube growth is associated with cancer-like metabolic reprogramming and is limited by phosphoglycerate dehydrogenase.
Article Snippet: 1 h before transfection, medium was changed to 7 ml of fresh growth medium (DMEM Glutamax (Gibco, Cat#10566016, Waltham, MA, USA), 10 % FBS), into which the transfection mixture was added. .. For transfection mixture, 4 μg of PHGDH plasmid or 4 μg of empty vector plasmid (Addgene, plasmid #52107) was combined with 4 μg of DNA RV helper plasmid (Addgene, plasmid #12371) in 1800 μl of Opti-MEM. .. 6 μl of Lipofectamine Plus reagent was then added and incubated for 5 min at room temperature, followed by 24 μl of Lipofectamine LTX (ThermoFisher, Cat#15338100, Waltham, MA, USA) for a further 30-min incubation at room temperature.
Article Title: SNCA -targeted epigenome therapy for Parkinson’s disease alleviates pathological and behavioral perturbations in a mouse model
Article Snippet: .. Briefly, 15 μg vector plasmid, 10 μg psPAX2 packaging plasmid (Addgene 12260 generated in Dr. Didier Trono’s lab, EPFL, Switzerland), 5 μg pMD2.G envelope plasmid (Addgene 12259, generated in Dr. Trono’s lab), and 2.5 μg pRSV-Rev plasmid (Addgene 12253, generated in Dr. Trono’s lab) were transfected into 293T cells. .. Vector particles were collected from filtered conditioned medium at 72 hr post-transfection.
Article Title: CRB2 Depletion Induces YAP Signaling and Disrupts Mechanosensing in Podocytes
Article Snippet: .. Briefly, 15 μg of vector plasmid, 10 μg of psPAX2 packaging plasmid (Addgene #12260 generated in Dr Didier Trono’s laboratory, EPFL, Switzerland), 5 μg of pMD2.G envelope plasmid (Addgene #12259, generated in Dr Trono’s laboratory) and 2.5 μg of pRSV-Rev plasmid (Addgene #12253, generated in Dr Trono’s laboratory) were transfected into 293 T cells. .. Vector particles were collected from the filtered conditioned medium at 72 h post-transfection.
Article Title: Neuronal-type-specific epigenome editing to decrease SNCA expression: Implications for precision medicine in synucleinopathies
Article Snippet: .. Briefly, 15mg of vector plasmid, 10mg of psPAX2 packaging plasmid (Addgene 12260), 5 mg of pMD2.G envelope plasmid (Addgene 12259), and 2.5 mg of pRSV-Rev plasmid (Addgene 12253)were transfected into 293T cells. .. Vector particles were collected from filtered conditioned medium at 72 h post transfection.
Article Title: A proof of concept gene-activated titanium surface for oral implantology applications.
Article Snippet: .. A plasmid encoding EGFP (Plasmid #13031, Addgene, UK), a plasmid encoding mouse PDGFB (RefSeq BC064056, Origene, Rockville, MD, USA), and an empty vector plasmid (Plasmid #52535, Addgene) were purified from DH5α Escherichia coli that had been previously transformed. .. Purification was performed using a GenElute HP EndotoxinFree Plasmid Maxiprep Kit (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer's protocol.
Generated:Article Title: CRB2 depletion induces YAP signaling and disrupts mechanosensing in podocytes.
Article Snippet: .. 160 Briefly, 15 μg of vector plasmid, 10 μg of psPAX2 packaging plasmid (Addgene #12260 161 generated in Dr Didier Trono’s laboratory, EPFL, Switzerland), 5 μg of pMD2.G envelope 162 plasmid (Addgene #12259, generated in Dr Trono’s laboratory) and 2.5 μg of pRSV-Rev 163 plasmid (Addgene #12253, generated in Dr Trono’s laboratory) were transfected into 293 T 164 cells. .. Vector particles were collected from the filtered conditioned medium at 72 h post-165 transfection.
Article Title: SNCA -targeted epigenome therapy for Parkinson’s disease alleviates pathological and behavioral perturbations in a mouse model
Article Snippet: .. Briefly, 15 μg vector plasmid, 10 μg psPAX2 packaging plasmid (Addgene 12260 generated in Dr. Didier Trono’s lab, EPFL, Switzerland), 5 μg pMD2.G envelope plasmid (Addgene 12259, generated in Dr. Trono’s lab), and 2.5 μg pRSV-Rev plasmid (Addgene 12253, generated in Dr. Trono’s lab) were transfected into 293T cells. .. Vector particles were collected from filtered conditioned medium at 72 hr post-transfection.
Article Title: CRB2 Depletion Induces YAP Signaling and Disrupts Mechanosensing in Podocytes
Article Snippet: .. Briefly, 15 μg of vector plasmid, 10 μg of psPAX2 packaging plasmid (Addgene #12260 generated in Dr Didier Trono’s laboratory, EPFL, Switzerland), 5 μg of pMD2.G envelope plasmid (Addgene #12259, generated in Dr Trono’s laboratory) and 2.5 μg of pRSV-Rev plasmid (Addgene #12253, generated in Dr Trono’s laboratory) were transfected into 293 T cells. .. Vector particles were collected from the filtered conditioned medium at 72 h post-transfection.
Transfection:Article Title: CRB2 depletion induces YAP signaling and disrupts mechanosensing in podocytes.
Article Snippet: .. 160 Briefly, 15 μg of vector plasmid, 10 μg of psPAX2 packaging plasmid (Addgene #12260 161 generated in Dr Didier Trono’s laboratory, EPFL, Switzerland), 5 μg of pMD2.G envelope 162 plasmid (Addgene #12259, generated in Dr Trono’s laboratory) and 2.5 μg of pRSV-Rev 163 plasmid (Addgene #12253, generated in Dr Trono’s laboratory) were transfected into 293 T 164 cells. .. Vector particles were collected from the filtered conditioned medium at 72 h post-165 transfection.
Article Title: Myotube growth is associated with cancer-like metabolic reprogramming and is limited by phosphoglycerate dehydrogenase.
Article Snippet: 1 h before transfection, medium was changed to 7 ml of fresh growth medium (DMEM Glutamax (Gibco, Cat#10566016, Waltham, MA, USA), 10 % FBS), into which the transfection mixture was added. .. For transfection mixture, 4 μg of PHGDH plasmid or 4 μg of empty vector plasmid (Addgene, plasmid #52107) was combined with 4 μg of DNA RV helper plasmid (Addgene, plasmid #12371) in 1800 μl of Opti-MEM. .. 6 μl of Lipofectamine Plus reagent was then added and incubated for 5 min at room temperature, followed by 24 μl of Lipofectamine LTX (ThermoFisher, Cat#15338100, Waltham, MA, USA) for a further 30-min incubation at room temperature.
Article Title: SNCA -targeted epigenome therapy for Parkinson’s disease alleviates pathological and behavioral perturbations in a mouse model
Article Snippet: .. Briefly, 15 μg vector plasmid, 10 μg psPAX2 packaging plasmid (Addgene 12260 generated in Dr. Didier Trono’s lab, EPFL, Switzerland), 5 μg pMD2.G envelope plasmid (Addgene 12259, generated in Dr. Trono’s lab), and 2.5 μg pRSV-Rev plasmid (Addgene 12253, generated in Dr. Trono’s lab) were transfected into 293T cells. .. Vector particles were collected from filtered conditioned medium at 72 hr post-transfection.
Article Title: CRB2 Depletion Induces YAP Signaling and Disrupts Mechanosensing in Podocytes
Article Snippet: .. Briefly, 15 μg of vector plasmid, 10 μg of psPAX2 packaging plasmid (Addgene #12260 generated in Dr Didier Trono’s laboratory, EPFL, Switzerland), 5 μg of pMD2.G envelope plasmid (Addgene #12259, generated in Dr Trono’s laboratory) and 2.5 μg of pRSV-Rev plasmid (Addgene #12253, generated in Dr Trono’s laboratory) were transfected into 293 T cells. .. Vector particles were collected from the filtered conditioned medium at 72 h post-transfection.
Article Title: Neuronal-type-specific epigenome editing to decrease SNCA expression: Implications for precision medicine in synucleinopathies
Article Snippet: .. Briefly, 15mg of vector plasmid, 10mg of psPAX2 packaging plasmid (Addgene 12260), 5 mg of pMD2.G envelope plasmid (Addgene 12259), and 2.5 mg of pRSV-Rev plasmid (Addgene 12253)were transfected into 293T cells. .. Vector particles were collected from filtered conditioned medium at 72 h post transfection.
other:Article Title: Human embryo implantation involves Syncytin-2/MFSD2A-mediated heterokaryon formation with maternal endometrium
Article Snippet: One week after electroporation edited organoids were selected with addition of Hygromycin B Gold (100 μg/ml; Invivogen) to the medium.
Purification:Article Title: A proof of concept gene-activated titanium surface for oral implantology applications.
Article Snippet: .. A plasmid encoding EGFP (Plasmid #13031, Addgene, UK), a plasmid encoding mouse PDGFB (RefSeq BC064056, Origene, Rockville, MD, USA), and an empty vector plasmid (Plasmid #52535, Addgene) were purified from DH5α Escherichia coli that had been previously transformed. .. Purification was performed using a GenElute HP EndotoxinFree Plasmid Maxiprep Kit (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer's protocol.
Transformation Assay:Article Title: A proof of concept gene-activated titanium surface for oral implantology applications.
Article Snippet: .. A plasmid encoding EGFP (Plasmid #13031, Addgene, UK), a plasmid encoding mouse PDGFB (RefSeq BC064056, Origene, Rockville, MD, USA), and an empty vector plasmid (Plasmid #52535, Addgene) were purified from DH5α Escherichia coli that had been previously transformed. .. Purification was performed using a GenElute HP EndotoxinFree Plasmid Maxiprep Kit (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer's protocol.
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