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Mission plko 1 puro non target shrna control plasmid dna

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The MISSION® pLKO.1-puro Non-Target shRNA Control Plasmid DNA is a laboratory tool used in RNA interference (RNAi) studies. It is a plasmid vector designed to express a non-targeting short hairpin RNA (shRNA) sequence, which serves as a control for RNAi experiments.

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Lab products found in correlation

X tremegene 9 dna transfection reagent, by Roche (3 mentions) Hexadimethrine bromide, by Merck Group (3 mentions) Hek293t cell, by American Type Culture Collection (2 mentions) Shc016, by Merck Group (2 mentions) Trcn0000020239, by Merck Group (2 mentions) Plko vector, by Thermo Fisher Scientific (1 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions) Pmd2 g, by Addgene (1 mentions)

Spelling variants of this product

PLKO.1-puro (99 citations) MISSION® shRNA Plasmid DNA (24 citations) Non-target shRNA control (11 citations) MISSION pLKO.1-puro (10 citations) PLKO.1-puro Non-Target shRNA Control Plasmid DNA (5 citations) PLKO.1-puro Non-Target shRNA (4 citations) MISSION pLKO.1-puro Non-Target shRNA Control Plasmid (2 citations) Mission® pLKO.1 puro non-target shRNA control (2 citations) Mission pLKO.1-puro non-target shRNA (2 citations) Control ShRNA pLKO.1 (2 citations)

4 protocols using mission plko 1 puro non target shrna control plasmid dna

1

Generating Stable SNAT2 Knockdown in Corneal Cells

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Plasmid expressing shRNA against human SNAT2 (Sigma-Aldrich, TRCN0000020239 and TRCN0000020241) and MISSION® pLKO.1-puro Non-Target shRNA Control Plasmid DNA (Sigma-Aldrich, SHC016)) were purchased from Sigma-Aldrich. HEK 293T cells (150 mm dishes, 50% confluency) (ATCC, CRL-3216) were transfected with 12.5 μg shRNA expressing vector: 8 μg viral packaging plasmid (psPAX2 (Addgene,12260)): 4.5 μg viral envelope plasmid (pMD2.G (Addgene,12259)) ratio with X-tremeGENE™ 9 DNA Transfection Reagent (Roche, 6365779001) according to the manufacturer’s instructions. After transfection, cells were maintained in corneal growth media which was collected after 24, 48 and 72 h and filtered through 0.45 μM filter. The filtrate was diluted with the corneal growth media (4:1 ratio) supplemented with Hexadimethrine bromide (Sigma-Aldrich, H9268) at 10μg/mL and added to the corneal cells for 12 h followed by 12 h in the growth media without viral particles. The procedure was repeated once more and cells were allowed to recover one day in the growth media. After this, cells were subcultured and grown in media containing 1 μg/mL puromycin. After 3–4 days of the selection with puromycin cells were used for experiments. Knockdown efficiency was verified by Western blot analysis.
Krokowski D., Jobava R., Szkop K.J., Chen C.W., Fu X., Venus S., Guan B.J., Wu J., Gao Z., Banaszuk W., Tchorzewski M., Mu T., Ropelewski P., Merrick W.C., Mao Y., Sevval A.I., Miranda H., Qian S.B., Manifava M., Ktistakis N.T., Vourekas A., Jankowsky E., Topisirovic I., Larsson O, & Hatzoglou M. (2022). Stress-induced perturbations in intracellular amino acids reprogram mRNA translation in osmoadaptation independently of the ISR. Cell reports, 40(3), 111092.
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2

Megalin Mini Receptor Plasmid Constructs

Cited 2 times
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Megalin mini receptor (mMeg) was generated from a human kidney cDNA library (Marzolo et al., 2003 (link); Farfán et al., 2013 (link)). Plasmids for phosphomimetic mMeg (mMeg S170D), non-phosphorylatable mMeg (mMeg S170A) and mMeg lacking the ectodomain (Meg0) were described previously (Yuseff et al., 2007 (link); Marzolo and Farfán, 2011 (link)). mCherry-Rab11 was kindly provided by Dr. Alexis Gautreau (Derivery et al., 2009 (link)). Human OCRL1-EGFP was described before (Vicinanza et al., 2011 (link)). Plasmids for short hairpin RNAs were MISSION® pLKO.1-puro Non-Target shRNA Control Plasmid DNA (Sigma-Aldrich), pLKO vectors purchased from Open Biosystems (shOCRL1 5′- GCC​AAG​TAT​AAG​AAA​GTT​CAA -3′ and shAPPL1 5′- GCA​TTG​TTA​GAA​CCT​CTA​CTT-3′). The primers used in quantitative PCR reactions were as follows: megalin forward, 5′- CTG​CTC​TTG​TAG​ACC​TGG​GTT​C -' 3; megalin reverse, 5′- TCG​GCA​CAG​CTA​CAC​TCA​TAA​C -3; glyceraldehyde-3-phosphate dehydrogenase forward, 5′- TCA​AGG​CTG​AGA​ATG​GGA​AG -`3; glyceraldehyde-3-phosphate dehydrogenase reverse, 5′- AGC​AGA​AGG​GGC​AGA​GAT​G -`3.
Sandoval L., Fuentealba L.M, & Marzolo M.P. (2022). Participation of OCRL1, and APPL1, in the expression, proteolysis, phosphorylation and endosomal trafficking of megalin: Implications for Lowe Syndrome. Frontiers in Cell and Developmental Biology, 10, 911664.
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3

Generating Stable SNAT2 Knockdown in Corneal Cells

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Plasmid expressing shRNA against human SNAT2 (Sigma-Aldrich, TRCN0000020239 and TRCN0000020241) and MISSION® pLKO.1-puro Non-Target shRNA Control Plasmid DNA (Sigma-Aldrich, SHC016)) were purchased from Sigma-Aldrich. HEK 293T cells (150 mm dishes, 50% confluency) (ATCC, CRL-3216) were transfected with 12.5 μg shRNA expressing vector: 8 μg viral packaging plasmid (psPAX2 (Addgene,12260)): 4.5 μg viral envelope plasmid (pMD2.G (Addgene,12259)) ratio with X-tremeGENE™ 9 DNA Transfection Reagent (Roche, 6365779001) according to the manufacturer’s instructions. After transfection, cells were maintained in corneal growth media which was collected after 24, 48 and 72 h and filtered through 0.45 μM filter. The filtrate was diluted with the corneal growth media (4:1 ratio) supplemented with Hexadimethrine bromide (Sigma-Aldrich, H9268) at 10μg/mL and added to the corneal cells for 12 h followed by 12 h in the growth media without viral particles. The procedure was repeated once more and cells were allowed to recover one day in the growth media. After this, cells were subcultured and grown in media containing 1 μg/mL puromycin. After 3–4 days of the selection with puromycin cells were used for experiments. Knockdown efficiency was verified by Western blot analysis.
Krokowski D., Jobava R., Szkop K.J., Chen C.W., Fu X., Venus S., Guan B.J., Wu J., Gao Z., Banaszuk W., Tchorzewski M., Mu T., Ropelewski P., Merrick W.C., Mao Y., Sevval A.I., Miranda H., Qian S.B., Manifava M., Ktistakis N.T., Vourekas A., Jankowsky E., Topisirovic I., Larsson O, & Hatzoglou M. (2022). Stress-induced perturbations in intracellular amino acids reprogram mRNA translation in osmoadaptation independently of the ISR. Cell reports, 40(3), 111092.
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4

Lentivirus-Mediated Knockdown of OPA1 in MEFs

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Plasmid expressing shRNA against OPA1 (Sigma-Aldrich, TRCN0000091111) and MISSION® pLKO.1-puro Non-Target shRNA Control Plasmid DNA (Sigma-Aldrich, SHC016)) were purchased from Sigma-Aldrich. HEK 293T cells (150 mm dishes, 50% confluency) (ATCC, CRL-3216) were transfected with 12.5 μg shRNA expressing vector: 8 μg viral packaging plasmid (psPAX2 (Addgene,12260)): 4.5 μg viral envelope plasmid (pMD2.G (Addgene,12259)) ratio with X-tremeGENE™ 9 DNA Transfection Reagent (Roche, 6365779001) according to the manufacturer’s instructions. Media was collected after 24, 48 and 72 hours and filtered through 0.45 μM filter. The filtrate was diluted with the full media (1:5 ratio) supplemented with Hexadimethrine bromide (Sigma-Aldrich, H9268) at 10μg/ml and added to MEFs for 12 h followed by 12 h in the growth media without viral particles. The procedure was repeated once more and cells were allowed to recover one day in the growth media. After this, cells were subcultured and grown in media containing 30 μg/ml puromycin (Thermo Fisher Scientific, A1113803). After 3–4 days of the selection with puromycin cells were used for experiments. Knockdown efficiency was verified using Western blot analysis.
Jobava R., Mao Y., Guan B.J., Hu D., Krokowski D., Chen C.W., Shu X.E., Chukwurah E., Wu J., Gao Z., Zagore L.L., Merrick W.C., Trifunovic A., Hsieh A.C., Valadkhan S., Zhang Y., Qi X., Jankowsky E., Topisirovic I., Licatalosi D.D., Qian S.B, & Hatzoglou M. (2021). Adaptive translational pausing is a hallmark of the cellular response to severe environmental stress. Molecular cell, 81(20), 4191-4208.e8.
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