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Pcdh ef1 t2a puro vector

Manufactured by System Biosciences
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The PCDH-EF1-T2A-Puro vector is a plasmid used for gene expression in mammalian cells. It contains a PCDH promoter, an EF1 constitutive promoter, a T2A self-cleaving peptide, and a puromycin resistance cassette.

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Lab products found in correlation

Peg it, by System Biosciences (3 mentions) In fusion hd cloning kit, by Takara Bio (2 mentions) Lenti x 293t cells, by Takara Bio (1 mentions)

Close spelling variants of this product

PCDH-EF1-MCS-BGH-PGK-GFP-T2A-Puro vector PCDH-EF1-MCS-T2A-Puro vector PCDH-CMV-MCS-EF1-GFP-T2A-Puro vector PCDH vector

3 protocols using pcdh ef1 t2a puro vector

1

Lentiviral Vector Production Protocol

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CAR constructs were either synthesized as gBlocks (IDT) or amplified by PCR from existing templates, and cloned into the EcoRI and NotI restriction sites of the pCDH-EF1-T2A-Puro vector (System Biosciences). All plasmid constructions were performed using the In-Fusion HD cloning kit (Clontech) and verified by DNA sequencing. For production of lentivirus, low-passage Lenti-X 293T cells (Clontech) were transfected with DNA using the PEI Max 40K (polyethyleneimine) technique, as described (8 (link)). Supernatant from Lenti-X 293T cell cultures was collected 2 days after transfection and passed through a 0.45 μM filter. Culture media were used for transduction either directly or concentrated using PEG-it (System Biosciences).
Zhuang X, & Long E.O. (2022). NK Cells Equipped With a Chimeric Antigen Receptor That Overcomes Inhibition by HLA Class I for Adoptive Transfer of CAR-NK Cells. Frontiers in Immunology, 13, 840844.
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2

Overexpressing and Silencing GS in Cells

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A GS-overexpressing lentiviral vector was constructed by cloning a full-length coding DNA fragment into the NheI/NotI sites of a pCDH-EF1-T2A-Puro vector (System Biosciences, Mountain View, CA, USA). Lentiviral-based short hairpin RNAs (shRNAs) targeting human GS (NM_002065.3-802s1c1, NM_002065.3-1450s1c1 and NM_002065.4-1890s21c1) and control shRNA targeting enhanced green fluorescent protein (GFP) (cat. no. SHC005V) were used for knockdown experiments.
Sohn B.H., Park I.Y., Shin J.H., Yim S.Y, & Lee J.S. (2018). Glutamine synthetase mediates sorafenib sensitivity in β-catenin-active hepatocellular carcinoma cells. Experimental & Molecular Medicine, 50(1), e421-.
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3

Generation of B7H7 and CD28H Constructs

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A plasmid containing B7H7 cDNA was obtained from Harvard PlasmID Database (#HsCD00044662). B7H7 cDNA was amplified and cloned into the EcoRI and NotI cloning sites of pAc5.1/V5-His A vector (Thermo Fisher Scientific) for expression in Drosophila S2 cells, and the EcoRI and NotI cloning sites of pCDH-EF1-T2A-Puro vector (System Biosciences) for expression in human cell lines. The cDNA of CD28H was obtained from Harvard PlasmID Database (#HsCD00416184) in the vector pLX304. CD28H cDNA was amplified and cloned into the EcoRI and NotI cloning sites of pCDH-EF1-T2A-Puro lentivirus vector (System Biosciences) for transduction of human cell lines. CD28H mutants and chimeras were generated using the In-Fusion HD cloning kit (Clontech) and verified by sequencing. All of the cDNAs cloned into the PCDH vector were in frame with the 2A-peptide. Expressed proteins could be detected by anti-2A antibody in immunoblots. All plasmid constructions were carried out using the In-Fusion HD cloning kit (Clontech).
Zhuang X, & Long E.O. (2019). CD28 homolog is a strong activator of natural killer cells for lysis of B7H7+ tumor cells. Cancer immunology research, 7(6), 939-951.
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