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Plko 1 backbone

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The PLKO.1 backbone is a plasmid vector commonly used in molecular biology and genetic engineering applications. It serves as a foundation for the construction of expression vectors, enabling the expression of genes of interest in various cell lines. The PLKO.1 backbone provides the necessary elements for plasmid replication and selection, facilitating the cloning and manipulation of genetic material.

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Lab products found in correlation

Shc003 turbo gfp, by Merck Group (2 mentions) Scramble shrna shc002, by Merck Group (2 mentions) Shc003, by Merck Group (2 mentions) Mission shrna lentiviral transduction, by Merck Group (2 mentions) Shrna lentiviral vector, by Merck Group (1 mentions) Pspax2, by Addgene (1 mentions) Pmd2 g, by Addgene (1 mentions)

Close spelling variants of this product

PLKO.1-puro backbone (2 citations) PLKO backbone (2 citations) PLKO.1 (97 citations)

6 protocols using plko 1 backbone

1

Cloning FAT1 and ZNF750 ORFs

Cited 2 times
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The open reading frame (ORF) of human FAT1 and ZNF750 transcripts were generously provided by Timothy Chan's group from Memorial Sloan-Kettering Cancer Center27 (link) and Paul Khavari's group from Veterans Affairs Palo Alto Healthcare System24 (link), respectively. Both ORFs were sub-cloned into lentiviral based expression vector SHC003 (Sigma-Aldrich) using Nhe I and Fse I cloning sites. SHC003-Turbo-GFP was used as control (Sigma-Aldrich). The lentiviral based shRNA vectors (shXPO1 and shFAT2 sequences are listed in Supplementary Table 13b) were generated with PLKO.1 backbone (Sigma-Aldrich) using Age I and EcoR I cloning sites. SHC002-Scramble shRNA was used as control (Sigma-Aldrich). The cloning primers are listed in Supplementary Table 13c.
Lin D.C., Hao J.J., Nagata Y., Xu L., Shang L., Meng X., Sato Y., Okuno Y., Varela A.M., Ding L.W., Garg M., Liu L.Z., Yang H., Yin D., Shi Z.Z., Jiang Y.Y., Gu W.Y., Gong T., Zhang Y., Xu X., Kalid O., Shacham S., Ogawa S., Wang M.R, & Koeffler H.P. (2014). Genomic and molecular characterization of esophageal squamous cell carcinoma. Nature genetics, 46(5), 467-473.
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2

Genetic Manipulation of ZNF750 in Cells

Cited 1 time
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Expression vectors pLEX-ZNF750 (wildtype) and pLEX-C2H2 mutant ZNF750 (C2H2 mutant) were generously provided by Dr Paul A. Khavari (Stanford University School of Medicine). We also used another ZNF750 expression vector, SHC003-ZNF750 (which we reported previously3 (link)), in some of the experiments to confirm that the results were not affected by different vector systems (Supplementary Figure 6). The shRNA construct for p63 was made with PLKO.1 backbone (Sigma-Aldrich) using Age I and EcoR I sites (Supplementary Table S3). LAMC2 over-expression and shRNA vectors were described previously.24 Pooled siRNAs targeting human ZNF750 (M-014417-01) and TINCR (R-015725-00-0005) and scramble siRNA (D-001210-01) were purchased from Thermo Scientific (Rockford, IL, USA) (Supplementary Table S3).
Hazawa M., Lin D.C., Handral H., Xu L., Chen Y., Jiang Y.Y., Mayakonda A., Ding L.W., Meng X., Sharma A., Samuel S., Movahednia M.M., Wong R.W., Yang H., Tong C, & Koeffler H.P. (2016). ZNF750 is a lineage-specific tumour suppressor in squamous cell carcinoma. Oncogene, 36(16), 2243-2254.
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3

Lentiviral shRNA Transfection Protocol

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All shRNA constructs were in the pLKO.1 backbone (Sigma). Transfection with indicated shRNA (Supplementary Table 1) and control empty shRNA (#SHC001) from Sigma were performed using FuGENE® HD Transfection Reagent (Roche Applied Science) or MISSION shRNA Lentiviral Transduction (Sigma) according to the manufacturer’s instructions.
Yang L., Xie M., Yang M., Yu Y., Zhu S., Hou W., Kang R., Lotze M., Billiar T.R., Wang H., Cao L, & Tang D. (2014). PKM2 Regulates the Warburg Effect and Promotes HMGB1 Release in Sepsis. Nature communications, 5, 4436.
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4

Lentiviral shRNA Transfection Protocol

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All shRNA constructs were in the pLKO.1 backbone (Sigma). Transfection with indicated shRNA (Supplementary Table 1) and control empty shRNA (#SHC001) from Sigma were performed using FuGENE® HD Transfection Reagent (Roche Applied Science) or MISSION shRNA Lentiviral Transduction (Sigma) according to the manufacturer’s instructions.
Yang L., Xie M., Yang M., Yu Y., Zhu S., Hou W., Kang R., Lotze M., Billiar T.R., Wang H., Cao L, & Tang D. (2014). PKM2 Regulates the Warburg Effect and Promotes HMGB1 Release in Sepsis. Nature communications, 5, 4436.
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5

Cloning FAT1 and ZNF750 ORFs

Cited 2 times
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The open reading frame (ORF) of human FAT1 and ZNF750 transcripts were generously provided by Timothy Chan's group from Memorial Sloan-Kettering Cancer Center27 (link) and Paul Khavari's group from Veterans Affairs Palo Alto Healthcare System24 (link), respectively. Both ORFs were sub-cloned into lentiviral based expression vector SHC003 (Sigma-Aldrich) using Nhe I and Fse I cloning sites. SHC003-Turbo-GFP was used as control (Sigma-Aldrich). The lentiviral based shRNA vectors (shXPO1 and shFAT2 sequences are listed in Supplementary Table 13b) were generated with PLKO.1 backbone (Sigma-Aldrich) using Age I and EcoR I cloning sites. SHC002-Scramble shRNA was used as control (Sigma-Aldrich). The cloning primers are listed in Supplementary Table 13c.
Lin D.C., Hao J.J., Nagata Y., Xu L., Shang L., Meng X., Sato Y., Okuno Y., Varela A.M., Ding L.W., Garg M., Liu L.Z., Yang H., Yin D., Shi Z.Z., Jiang Y.Y., Gu W.Y., Gong T., Zhang Y., Xu X., Kalid O., Shacham S., Ogawa S., Wang M.R, & Koeffler H.P. (2014). Genomic and molecular characterization of esophageal squamous cell carcinoma. Nature genetics, 46(5), 467-473.
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6

Polycistronic Construct pMXs-puro-MGT

Cited 1 time
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The polycistronic construct pMXs-puro-MGT was constructed as previously described [14 (link)]. The plasmid map of pMXs-puro-MGT is provided in Figure
S1. shRNA lentiviral vectors with pLKO.1 backbone were obtained from Sigma-Aldrich. Packaging and envelop vectors for lentivirus were psPAX2 and pMD2.G (Addgene).
Zhou Y., Alimohamadi S., Wang L., Liu Z., Wall J.B., Yin C., Liu J, & Qian L. (2018). A Loss of Function Screen of Epigenetic Modifiers and Splicing Factors during Early Stage of Cardiac Reprogramming. Stem Cells International, 2018, 3814747.
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