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.
Scrambled transduction particles plko 1 puro shrna vector only
Manufactured by Merck Group
The Scrambled transduction particles (pLKO.1-puro shRNA vector-only) is a lentiviral vector designed for the delivery of short hairpin RNA (shRNA) into target cells. The vector contains a puromycin resistance gene, allowing for the selection of transduced cells. This product is intended for use in research applications involving gene knockdown studies.
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2 protocols using scrambled transduction particles plko 1 puro shrna vector only
1
Silencing p38γ and PIK3CA in Hut78 and H9 cells
2
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.
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