Pclx ptf r1 dest r2 ebr65 lentiviral

Manufactured by Addgene

PCLX-pTF-R1-DEST-R2-EBR65 is a lentiviral vector that can be used for the expression of genes of interest in target cells. The vector contains the necessary elements for lentiviral packaging and transduction, as well as a Gateway cloning site for the insertion of your gene of interest.

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Lab products found in correlation

Pcmv6 an mrfp, by OriGene (2 mentions) Pcmv6 an myc ddk, by OriGene (2 mentions) Pegfp rnaseh1, by Addgene (2 mentions)

2 protocols using pclx ptf r1 dest r2 ebr65 lentiviral

Inducible GFP-RNAseH1 Plasmid Generation

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pCMV6-AN-Myc-DDK (PS100016; RRID:SCR_021264) and pCMV6-AN-mRFP (PS100049; RRID: SCR_021265) were purchased from Origene. Inducible GFP-RNAseH1 plasmid was generated via restriction cloning an RNH1-GFP cassette from pEGFP‐RNASEH1, a gift from Andrew Jackson and Martin Reijns (RRID: Addgene_108699) using BglII + NotI and cloning into the BamH1 NotI sites of pENTR1A (RRID:SCR_021263). The RNH1-GFP cassette was then transferred to the pCLX-pTF-R1-DEST-R2-EBR65 lentiviral plasmid, a gift from Patrick Salmon (RRID:Addgene_45952) via an LR (recombination reaction between attL and attR sites) recombination reaction, creating a single vector that expresses the Tet-repressor and blasticidin resistance gene under the control of a constitutive promoter and containing the RNHA1-GFP cassette under the control of a Tet-repressor regulated promoter. SF3B1-K700E and RNAseH1 D210N mutations were created by site-directed mutagenesis.

Lappin K.M., Barros E.M., Jhujh S.S., Irwin G.W., McMillan H., Liberante F.G., Latimer C., La Bonte M.J., Mills K.I., Harkin D.P., Stewart G.S, & Savage K.I. (2022). Cancer-Associated SF3B1 Mutations Confer a BRCA-Like Cellular Phenotype and Synthetic Lethality to PARP Inhibitors. Cancer Research, 82(5), 819-830.

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Generating Inducible GFP-RNAseH1 Plasmid

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pCMV6-AN-Myc-DDK (PS100016; RRID:SCR_021264) and pCMV6-AN-mRFP (PS100049; RRID: SCR_021265) were purchased from Origene. Inducible GFP-RNAseH1 plasmid was generated via restriction cloning an RNH1-GFP cassette from pEGFP-RNASEH1, a gift from Andrew Jackson & Martin Reijns (RRID:Addgene_108699) using BglII + NotI and cloning into the BamH1 NotI sites of pENTR1A (RRID:SCR_021263). The RNH1-GFP cassette was then transferred to the pCLX-pTF-R1-DEST-R2-EBR65 lentiviral plasmid, a gift from Patrick Salmon (RRID:Addgene_45952) via an LR recombination reaction, creating a single vector that expresses the Tet-repressor and blasticidin resistance gene under the control of a constitutive promoter and containing the RNHA1-GFP cassette under the control of a Tet-repressor regulated promoter. SF3B1-K700E and RNAseH1 D210N mutations where created by site-directed-mutagenesis.

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