Lenti x packaging single shots protocol

Manufactured by Takara Bio

The Lenti-X Packaging Single Shots Protocol is a laboratory product designed for the concentration and purification of lentiviral particles. It provides a streamlined workflow for the production of high-titer lentiviral stocks. This product includes all the necessary components and detailed instructions to efficiently concentrate and purify lentiviral particles from cell culture supernatants.

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Lab products found in correlation

Lenti x concentrator protocol, by Takara Bio (2 mentions) Plvx tetone puro, by Takara Bio (1 mentions) Plvx tetone, by Takara Bio (1 mentions)

Spelling variants of this product

Lenti-X (77 citations) Lenti-X Packaging Single Shots (39 citations) Lenti-x shots (3 citations)

2 protocols using lenti x packaging single shots protocol

Inducible PARG Expression System

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pLVX-TetOne-Puro (Clontech/Takara, 631,847) with puromycin selection was used as the backbone vector for an inducible gene expression system. Human PARG cDNA were cloned to pLVX-TetOne-Puro, followed by P2A cleavage site and mClover3 gene that encodes fluorescence protein with NLS signal for the visual control of the promoter activity.
Packaging Lenti-X Vectors into lentiviral particles and virus tittering: The lentiviral construct, pLVX-TetOne-hPARG-mClover3-NLS-Puro was packaged into lentiviral particles using Lenti-X Packaging Single Shots Protocol from Clontech/Takara. Lentiviral vector stocks were collected and concentrated using the Lenti-X Concentrator Protocol (Clontech/Takara). Lenti-X virus stocks were tittered using puromycin selection with HT1080 cells; the titer of virus corresponded to the number of colonies generated by the highest dilution multiplied by the dilution factor.
ccRCC cell lines were infected with lentivirus, selected with puromycin and single clones were generated using FACS sorting or with cloning rings. Clones with the highest fold change of hPARG induction vs. control detected with Western blotting were selected for propagation and further experiments.

Karpova Y., Guo D., Makhov P., Haines A.M., Markov D.A., Kolenko V, & Tulin A.V. (2021). Poly(ADP)-Ribosylation Inhibition: A Promising Approach for Clear Cell Renal Cell Carcinoma Therapy. Cancers, 13(19), 4973.

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Lentiviral Construct Packaging and Titration

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The lentiviral construct, pLVX-TetOne-hPARG-IRES-mClover3-NLS-Puro was packaged into lentiviral particles using Lenti-X Packaging Single Shots Protocol from Clontech/Takara. Lentiviral vector stocks were collected and concentrated using the Lenti-X Concentrator Protocol (Clontech/Takara). Lenti-X virus stocks were tittered using puromycin selection with HT1080 cells; the titer of virus corresponded to the number of colonies generated by the highest dilution multiplied by the dilution factor.

Johnson S., Karpova Y., Guo D., Ghatak A., Markov D.A, & Tulin A.V. (2022). PARG suppresses tumorigenesis and downregulates genes controlling angiogenesis, inflammatory response, and immune cell recruitment. BMC Cancer, 22, 557.

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