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Gsk lsd1

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GSK-LSD1 is a potent and selective inhibitor of the LSD1 (lysine-specific demethylase 1) enzyme. LSD1 is an epigenetic regulator that plays a role in the demethylation of mono- and di-methylated histone H3 at lysine 4 (H3K4). The GSK-LSD1 compound inhibits the catalytic activity of LSD1, leading to the accumulation of methylated histones and the modulation of gene expression.

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Lab products found in correlation

Propidium iodide, by Thermo Fisher Scientific (3 mentions) Ory 1001, by Selleck Chemicals (2 mentions) Stereotaxic frame, by Stoelting (2 mentions) Tnp470, by Cayman Chemical (1 mentions) Tranylcypromine, by Merck Group (1 mentions) Homocysteine, by Merck Group (1 mentions) Cerulenin, by Cayman Chemical (1 mentions) Hypoxanthine, by Merck Group (1 mentions) Apc cd11b antibody, by BioLegend (1 mentions) 6 thioguanine, by Cayman Chemical (1 mentions) 13c5 hypoxanthine, by Cambridge Isotopes (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)

4 protocols using gsk lsd1

1

Pharmacological and Viral Manipulation of LSD1 and EGR1 in Mouse Brain

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GSK-LSD1 (Tocris, Cat. # 5361) or ORY-1001 (SelleckChem, CAS # 1431326–61-2) was dissolved in water as a stock solution and then further diluted in sterile saline as working solution. Mice were injected with GSK-LSD1 (5 mg/kg, i.p.), ORY-1001 (0.015 mg/kg, i.p.) or vehicle control once daily for three consecutive days.
For viral injection, the mouse was deeply anesthetized with ketamine/xylazine (100mg/kg; 10mg/kg) and placed in a stereotaxic frame (Stoelting, USA). Under standard and sterile surgical conditions, a 10 μl syringe (7000 series, Hamilton, USA) attached to a micropump was lowered through to skull burr hole into the PFC (AP +1.8 mm, L ±0.3 mm, DV −2.7 mm). Mice were injected with either AAV2-CMV-GFP or AAV2-CMV-EGR1-Flag in the PFC (0.3 μl per hemisphere) at a flow rate of 50 nl/min.
Rapanelli M., Williams J.B., Ma K., Yang F., Zhong P., Patel R., Kumar M., Qin L., Rein B., Wang Z.J., Kassim B., Javidfar B., Couto L., Akbarian S, & Yan Z. (2022). Targeting Histone Demethylase LSD1 for Treatment of Deficits in Autism Mouse Models. Molecular psychiatry, 27(8), 3355-3366.
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2

Validation of Small-Molecule Inhibitors in iNeurons

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For validation using small-molecule inhibitors in iNeurons, SNCA-tagged CRISPRi-iPSCs were differentiated to iNeurons. Day 7 iNeurons were treated with the drugs, TNP470 (Cayman Chemical #16449) or GSK-LSD1 (Tocris #5361) dissolved in dimethyl sulfoxide (DMSO), or DMSO only (control) at different desired concentrations. The iNeurons were harvested using papain for FACS analysis on days 14 and 20. All experiments were performed in duplicate.
Santhosh Kumar S., Naseri N.N., Pather S.R., Hallacli E., Ndayisaba A., Buenaventura C., Acosta K., Roof J., Fazelinia H., Spruce L.A., Luk K., Khurana V., Rhoades E, & Shalem O. (2024). Sequential CRISPR screening reveals partial NatB inhibition as a strategy to mitigate alpha-synuclein levels in human neurons. Science Advances, 10(6), eadj4767.
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3

Pharmacological and Viral Manipulation of LSD1 and EGR1 in Mouse Brain

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GSK-LSD1 (Tocris, Cat. # 5361) or ORY-1001 (SelleckChem, CAS # 1431326–61-2) was dissolved in water as a stock solution and then further diluted in sterile saline as working solution. Mice were injected with GSK-LSD1 (5 mg/kg, i.p.), ORY-1001 (0.015 mg/kg, i.p.) or vehicle control once daily for three consecutive days.
For viral injection, the mouse was deeply anesthetized with ketamine/xylazine (100mg/kg; 10mg/kg) and placed in a stereotaxic frame (Stoelting, USA). Under standard and sterile surgical conditions, a 10 μl syringe (7000 series, Hamilton, USA) attached to a micropump was lowered through to skull burr hole into the PFC (AP +1.8 mm, L ±0.3 mm, DV −2.7 mm). Mice were injected with either AAV2-CMV-GFP or AAV2-CMV-EGR1-Flag in the PFC (0.3 μl per hemisphere) at a flow rate of 50 nl/min.
Rapanelli M., Williams J.B., Ma K., Yang F., Zhong P., Patel R., Kumar M., Qin L., Rein B., Wang Z.J., Kassim B., Javidfar B., Couto L., Akbarian S, & Yan Z. (2022). Targeting Histone Demethylase LSD1 for Treatment of Deficits in Autism Mouse Models. Molecular psychiatry, 27(8), 3355-3366.
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4

Non-Adherent Cell Drug Screening

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For non-screen drug plating, 2.5–5×103 live cells were seeded onto flat-bottom 96-well plastic plates in 200 μL of media per well. Drugs were added such that the total DMSO (Sigma-Aldrich) concentration was the same for all wells. After treatment, plates were transferred onto round-bottom 96-well plastic plates (Greiner Bio-One). Cells were analyzed on an iQue Screener Plus-VBR flow cytometer with a similar gating strategy as the one used for the screen, except we isolated singlets using both height and area parameters for FSC and SSC from live cells. We then isolated GFP + or antibody-positive events from the live singlets. GSK-LSD1, mercaptopurine (Tocris), cerulenin (Cayman Chemical), 6-thioguanine (Cayman Chemical), guanosine (Cayman Chemical), TOFA (MedChemExpress), ND-630 (MedChemExpress), hypoxanthine (Sigma Aldrich), and 13C5-hypoxanthine (Cambridge Isotope Laboratories) were dissolved in DMSO as 10mM stocks. Tranylcypromine (Sigma Aldrich) and homocysteine (Sigma Aldrich) were dissolved in ethanol and water respectively as 100mM stock. APC-CD11b antibody (BioLegend) and propidium iodide (ThermoFisher) were used at concentrations of 1 μg/mL or 50 μg/mL respectively.
Zee B.M., Poels K.E., Yao C.H., Kawabata K.C., Wu G., Duy C., Jacobus W.D., Senior E., Endress J.E., Jambhekar A., Lovitch S.B., Ma J., Dhall A., Harris I.S., Blanco M.A., Sykes D.B., Licht J.D., Weinstock D.M., Melnick A., Haigis M.C., Michor F, & Shi Y. (2021). Combined epigenetic and metabolic treatments overcome differentiation blockade in acute myeloid leukemia. iScience, 24(6), 102651.
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