Mission plko 1 puro non target shrna control transduction particles shc016v 1ea

Manufactured by Merck Group

The Mission pLKO.1-puro Non-target shRNA control transduction particles (SHC016V-1EA) are a laboratory tool used for gene expression studies. They are designed to produce shRNA that does not target any known gene, serving as a control for RNAi experiments. The core function of these particles is to facilitate the delivery and expression of a non-targeting shRNA sequence in cells.

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Lab products found in correlation

Mission shrna lentiviral transduction particles, by Merck Group (2 mentions) Puromycin, by Thermo Fisher Scientific (2 mentions)

3 protocols using mission plko 1 puro non target shrna control transduction particles shc016v 1ea

PHGDH knockdown in platinum-sensitive cells

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PHGDH was knocked-down in platinum sensitive cells using two commercially available shRNAs targeting PHGDH (TRCN0000255351 and TRCN0000233029), using MISSION shRNA lentiviral transduction particles (Sigma). Mission pLKO.1-puro Non-target shRNA control transduction particles (SHC016V-1EA, Sigma) were used as control. In total, 150,000 cells were seeded per well in a 12-well plate. The next day, the lentivirus was put on the cells with a MOI of 0.5 in the presence of 5 µg/ml polybrene. After 24 h, medium was replaced with pre-warmed complete growth medium. On day 3, infected cells were selected and further grown in the presence of 2 µg/ml puromycin (Gibco).

Van Nyen T., Planque M., van Wagensveld L., Duarte J.A., Zaal E.A., Talebi A., Rossi M., Körner P.R., Rizzotto L., Moens S., De Wispelaere W., Baiden-Amissah R.E., Sonke G.S., Horlings H.M., Eelen G., Berardi E., Swinnen J.V., Berkers C.R., Carmeliet P., Lambrechts D., Davidson B., Agami R., Fendt S.M., Annibali D, & Amant F. (2022). Serine metabolism remodeling after platinum-based chemotherapy identifies vulnerabilities in a subgroup of resistant ovarian cancers. Nature Communications, 13, 4578.

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Stable TET2 Knockdown in HCT116 Cells

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Stable knockdown of TET2 in HCT116 used MISSION shRNA Lentiviral Transduction Particles (SHCLNV-NM_017628, Sigma) following manufacturer’s instructions and selection with 2 ug/mL Puromycin (A11138-03, Life Technologies). For TET2C 10MOI of TRCN0000418976 and for TET2 + 3 5MOI each of TRCN0000418976 and TRCN0000246258 were used. shRNA control used 10MOI of MISSION pLKO.1-puro Non-Target shRNA Control Transduction Particles (SHC016V-1EA, Sigma).

Uribe-Lewis S., Stark R., Carroll T., Dunning M.J., Bachman M., Ito Y., Stojic L., Halim S., Vowler S.L., Lynch A.G., Delatte B., de Bony E.J., Colin L., Defrance M., Krueger F., Silva A.L., ten Hoopen R., Ibrahim A.E., Fuks F, & Murrell A. (2015). 5-hydroxymethylcytosine marks promoters in colon that resist DNA hypermethylation in cancer. Genome Biology, 16(1), 69.

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Stable PU.1 Overexpression and Knockdown in BV2 Cells

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Lenti ORF particles, Sfpi1 (Myc-DDK-tagged), pLenti-C-Myc-DDK-P2A-Puro (MR203632L3V) and LentiORF control particles of pLenti-C-Myc-DDK-P2A-Puro (PS100092V) were purchased from OriGene Technologies. Mouse SPI1 MISSION shRNA Lentiviral pLKO.1 Particles (TRCN0000009500) and MISSION® pLKO.1-puro Non-Target shRNA Control Transduction Particles (SHC016V-1EA) were purchased from Sigma-Aldrich. BV2 cells were seeded in 24-well plates at 50.000 per well and transduced the next day with lentiviral particles at MOI 5 using spinfection protocol 2000 rpm for 2 hours at room temperature (RT). After 48 hours of incubation, cells were split into a 6-well plate and selected with 10 μg/ml puromycin for 5 days. Surviving cells were amplified and frozen to create BV2 cells with overexpression of mouse PU.1 (OE) and respective empty frame control (CTL) or cells with knock-down of PU.1 (KD) with the respective scrambled control (SCR). Changes in PU.1 expression were confirmed on the mRNA level using qPCR and protein level using western blotting of polyclonal selected lines.

Pimenova A.A., Herbinet M., Gupta I., Machlovi S.I., Bowles K.R., Marcora E, & Goate A.M. (2020). Alzheimer’s-associated PU.1 expression levels regulate microglial inflammatory response. Neurobiology of disease, 148, 105217.

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