Plko 1 puro shrna vector

Manufactured by Merck Group

The PLKO.1-puro shRNA vector is a laboratory tool designed for the expression of short hairpin RNA (shRNA) molecules. It provides a platform for the delivery and stable expression of shRNA constructs in mammalian cell lines. The vector contains a puromycin resistance gene, allowing for the selection of successfully transfected cells.

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Lab products found in correlation

Scrambled transduction particles plko 1 puro shrna vector only, by Merck Group (2 mentions) Lenti shrna scramble control particles, by OriGene (2 mentions) Lentiviral transduction particles, by Merck Group (2 mentions) Polybrene, by Merck Group (2 mentions) Erastin e7781, by Selleck Chemicals (1 mentions) Ly2090314, by Selleck Chemicals (1 mentions) Fer 1 s7243, by Selleck Chemicals (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Mda mb 231, by American Type Culture Collection (1 mentions) Hek293t, by American Type Culture Collection (1 mentions)

3 protocols using plko 1 puro shrna vector

Genetic manipulation of GSK-3β in cell lines

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All cell lines (HeLa, MDA-MB-231, and HEK293T) were obtained from the American Type Culture Collection and were maintained in Dulbecco’s modified Eagle medium (DMEM, Gibco, 8121298) supplemented with 10% fetal bovine serum (FBS, Gibco, 2176398) and 1% penicillin–streptomycin at 37 °C with 5% CO₂.
The following regents were used in this study: Erastin (E7781), Fer-1 (S7243), and LY2090314 (S7063) were purchased from Selleck (Houston, TX, USA). The coding region of GSK-3β gene was cloned into the pCMV vector in-frame with a myc tag within the vector. The pLKO.1-puro shRNA vector was obtained from Sigma-Aldrich (SHC001). The pLKO.1-shGSK-3β 1^# and 2^# were constructed using specific primer containing separate targeting sequence. (targeting sequence to GSK-3β 1^#: 5′-CACTGGTCACGTTTGGAAAGA-3′, GSK-3β 2^#: 5′-CCCAAACTACACAGAATTTAA-3′).

Wang L., Ouyang S., Li B., Wu H, & Wang F. (2021). GSK-3β manipulates ferroptosis sensitivity by dominating iron homeostasis. Cell Death Discovery, 7, 334.

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Silencing p38γ and PIK3CA in Hut78 and H9 cells

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MISSION shRNA Lentiviral Transduction Particles (company validated) expressing five shRNAs in pLKO.1-puro shRNA vector that targets four exons of the human p38γ gene (MAPK12) and scrambled transduction particles (pLKO.1-puro shRNA vector-only) were purchased from Sigma-Aldrich. Four shRNA lentiviral particles each contain unique 29mer target human PIK3CA-specific shRNA (catalog #TL310428V) and one Lenti shRNA scramble control particles (catalog #TR30021V) are from OriGene Technologies, Inc. (Rockville, MD). Hut78 and H9 cells growing exponentially (70%–80% confluent) were seeded into 6-well plates (2 × 10⁵ cells/well) before transduction. Viral transduction efficiencies were improved by adding polybrene (hexadimethrine bromide, Sigma-Aldrich) to a final concentration of 8–10 μg/ml. Hut78 and H9 cells were transduced at a multiplicity of infection of 2. Hut78 and H9 cells were incubated overnight with the medium containing lentiviral particles and polybrene; then the medium containing lentiviral particles was removed from wells and replaced with fresh medium. Cells were collected on day 5 after transduction before RNA extraction and protein isolation.

Zhang X.H., Nam S., Wu J., Chen C.H., Liu X., Li H., McKeithan T., Gong Q., Chan W.C., Yin H.H., Yuan Y.C., Pillai R., Querfeld C., Horne D., Chen Y, & Rosen S.T. (2018). Multi-Kinase Inhibitor with Anti-p38γ Activity in Cutaneous T-Cell Lymphoma. The Journal of investigative dermatology, 138(11), 2377-2387.

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Silencing p38γ and PIK3CA in Hut78 and H9 cells

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