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Pcdh gfp

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The PCDH-GFP is a plasmid containing the Protocadherin (PCDH) gene fused to the Green Fluorescent Protein (GFP) gene. It is designed for the expression of the PCDH-GFP fusion protein in mammalian cell lines.

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Lab products found in correlation

Packaging vectors, by System Biosciences (2 mentions) Amicon ultra 15 centrifugal filter unit, by Merck Group (1 mentions) Pcmv dr8.2 dvpr, by Addgene (1 mentions) Polyethylenimine (pei), by Polysciences (1 mentions) Facsaria cell sorter, by BD (1 mentions) Polybrene, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

PCDH-MSCV-EF1-GFP-T2A-Pu PCDH-CuO-MCS-EF1-GFP lentiviral vector PCDH-EF1-MCS-T2A-GFP third generation vector PCDH-EF1α-MCS-T2A-GFP vector PCDH-CMV-MCS-EF1-GFP lentiviral vector PCDH-MSCVMCS-EF1α-GFP+Puro PCDH-CMV-MCS-EF1-GFP-T2A-Puro vector PCDH-CMV-MCS-EF1a-GFP-Neo PCDH-EF1a-MCS-BGH-PGK-GFP-T2A-Puro PCDH-MSCV-mcs-EF1-GFP-T2A-Pu

7 protocols using pcdh gfp

1

SARS-CoV-2 Pseudovirus Production and Infection

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HIV-GFP virus pseudotyped with SARS-CoV-2 S protein were generated by co-transfecting pcDNA3.1-SARS-CoV-2-S (a gift from Dr. Lu Lu at Fudan University), pCMV-dR8.2 dvpr (Addgene, 8455) (encoding HIV backbone) and pCDH-GFP (System Bioscience, CD-511B-1) (encoding the GFP reporter) at 1:2:3 mass ratio into Expi293F cells using PEI (Polysciences) at 3:1 (PEI:DNA) mass ratio. Four days post-transfection, the supernatant was harvested and filtered through a 0.45 μm sterile syringe filter, followed by concentration using Amicon Ultra-15 Centrifugal Filter Unit (Millipore, UFC910024). Pseudoviruses were titrated by HIV-1 p24 ELISA Assay (XpressBio) according to the manufacturer’s protocol and stored at −80°C. Cells were co-cultured with the pseudovirus (p24 = ∼60 ng/mL) for 12 hr and resuspended in fresh media, followed by an additional 36 hr incubation before analysis by flow cytometry to determine the GFP expression.
Lu Q., Liu J., Zhao S., Gomez Castro M.F., Laurent-Rolle M., Dong J., Ran X., Damani-Yokota P., Tang H., Karakousi T., Son J., Kaczmarek M.E., Zhang Z., Yeung S.T., McCune B.T., Chen R.E., Tang F., Ren X., Chen X., Hsu J.C., Teplova M., Huang B., Deng H., Long Z., Mudianto T., Jin S., Lin P., Du J., Zang R., Su T.T., Herrera A., Zhou M., Yan R., Cui J., Zhu J., Zhou Q., Wang T., Ma J., Koralov S.B., Zhang Z., Aifantis I., Segal L.N., Diamond M.S., Khanna K.M., Stapleford K.A., Cresswell P., Liu Y., Ding S., Xie Q, & Wang J. (2021). SARS-CoV-2 exacerbates proinflammatory responses in myeloid cells through C-type lectin receptors and Tweety family member 2. Immunity, 54(6), 1304-1319.e9.
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2

Generation of TROY-overexpressing NF-κB reporter cell lines

Cited 2 times
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A cDNA fragment encoding WT TROY with a C-terminal 3X HA epitope-tag was subcloned into the lentiviral transfer vector pCDH GFP (System Biosciences) as previously described [14] (link). The empty pCDH lentiviral construct expressing only GFP was used as control. Recombinant lentiviruses encoding TROY-HA were produced by transient transfection of 293T packing cells with pCDH construct encoding HA tagged TROY and the pPACKH1 plasmid packing mix (System Biosciences). 293/NF-κB-luc cells, a NF-κB response element–driven firefly luciferase reporter cell line [15] (link), and T98G/NF-κB-luc cells were transduced with the recombinant TROY-HA lentiviruses and selected by mass sorting the GFP-positive cells on a FACS Aria cell sorter (BD Biosciences) to generate the reporter cell lines designated 293/NF-κB-luc/TROY-HA and T98G/NF-κB-luc/TROY-HA, respectively.
Ding Z., Dhruv H., Kwiatkowska-Piwowarczyk A., Ruggieri R., Kloss J., Symons M., Pirrotte P., Eschbacher J.M., Tran N.L, & Loftus J.C. (2018). PDZ-RhoGEF Is a Signaling Effector for TROY-Induced Glioblastoma Cell Invasion and Survival. Neoplasia (New York, N.Y.), 20(10), 1045-1058.
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3

Expressing APLNR Protein in Mammalian Cells

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pcDNA3.1+-DYK-human APLNR (OHu11900) expressing the human APLNR protein with an N-terminal DYKDDDDK (FLAG) tag in mammalian cells (pcDNA-APLNR) was purchased from GenScript Biotech Corporation (Piscataway, NJ). An empty vector, pCDH-CMV-MCS-EF1-GreenPuro (CD513B-1; pCDH-GFP), was purchased from System Bioscience (CA, USA), and the FLAG-tagged APLNR gene from the abovementioned plasmid was subcloned (pCDH-GFP-APLNR). Mammalian cells transfected with this plasmid express APLNR and GFP under the control of the CMV and EF1 promoters, respectively.
Kinjo T., Ebisawa S., Nokubo T., Hashimoto M., Yamada T., Oshio M., Nakamura R., Uno K, & Kuramoto N. (2023). Post-translational modifications of the apelin receptor regulate its functional expression. AIMS Neuroscience, 10(4), 282-299.
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4

Lentiviral Transduction of RIP140 in Neurons

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Full length RIP140 or shRIP140 (target sequence: 5’-CCCGGCGTTGACATCAAAGAA-3’ and 5’-AAGCTTCTTTCTTTAA-3’) was subcloned into the site of NotI and XbaI of pCDH-GFP (System Biosciences) to generate RIP140 overexpression or RIP140 silence lentiviral vectors. Lentivirus was concentrated and titled as described63 (link). Briefly, 293T cells were transfected with expression vectors (full length RIP140 or shRIP140) and packaging vectors (System Biosciences). Lentivirus-containing medium was collected at 24, 48 and 72 h after transfection. Viral particles were concentrated by ultracentrifugation and purified through a sucrose cushion (20% sucrose in HBSS). Concentrated and purified virus was suspended in HBSS and stored at −80°C. For transduction, neurons were incubated with lentivirus for 16 h and then the medium was replaced with fresh culture medium.
Feng X., Krogh K.A., Wu C.Y., Lin Y.W., Tsai H.C., Thayer S.A, & Wei L.N. (2014). Receptor interacting protein 140 attenuates endoplasmic reticulum stress in neurons and protects against cell death. Nature communications, 5, 4487.
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5

Lentiviral Transduction of RIP140 in Neurons

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Full length RIP140 or shRIP140 (target sequence: 5’-CCCGGCGTTGACATCAAAGAA-3’ and 5’-AAGCTTCTTTCTTTAA-3’) was subcloned into the site of NotI and XbaI of pCDH-GFP (System Biosciences) to generate RIP140 overexpression or RIP140 silence lentiviral vectors. Lentivirus was concentrated and titled as described63 (link). Briefly, 293T cells were transfected with expression vectors (full length RIP140 or shRIP140) and packaging vectors (System Biosciences). Lentivirus-containing medium was collected at 24, 48 and 72 h after transfection. Viral particles were concentrated by ultracentrifugation and purified through a sucrose cushion (20% sucrose in HBSS). Concentrated and purified virus was suspended in HBSS and stored at −80°C. For transduction, neurons were incubated with lentivirus for 16 h and then the medium was replaced with fresh culture medium.
Feng X., Krogh K.A., Wu C.Y., Lin Y.W., Tsai H.C., Thayer S.A, & Wei L.N. (2014). Receptor interacting protein 140 attenuates endoplasmic reticulum stress in neurons and protects against cell death. Nature communications, 5, 4487.
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6

Generating Stable HIF1A-As2 Expression

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Full-length and truncated constructs of HIF1A-As2 were PCR amplified and inserted into a lentiviral vector (pCDH-GFP, System Biosciences). The inserted sequences were examined by sequencing. To produce lentiviruses with HIF1A-As2 stable expression, HEK293 cells were transfected with pCDH-HIF1A-As2 plasmid and package plasmids (Gag:pol, VSVG and REV) using Lipofectamine 2000 regent. Infectious lentiviruses were collected at 48, 72 and 96 h after transfection and filtered through 0.45 μm filters. H1299 cells were infected with the virus in the presence of polybrene (Santa Cruz) for 48 h and were sorted based on GFP expression using Flow Cytometry (Novocyte). Primers are listed in Supplementary Table 2.
Yang K., Zhang W., Zhong L., Xiao Y., Sahoo S., Fassan M., Zeng K., Magee P., Garofalo M, & Shi L. (2023). Long non-coding RNA HIF1A-As2 and MYC form a double-positive feedback loop to promote cell proliferation and metastasis in KRAS-driven non-small cell lung cancer. Cell Death and Differentiation, 30(6), 1533-1549.
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7

Lentiviral Transduction of KIMAT1 Constructs

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KIMAT1 full-length or deletion constructs were PCR amplified and inserted into a lentiviral vector (pCDH-GFP, System Biosciences). The inserted sequences were validated by sequencing. Cells were sorted based on GFP expression using Flow Cytometry (Novocyte NovoExpress Software, version 1.3.0). Primers are listed in Supplementary Table 3.
Shi L., Magee P., Fassan M., Sahoo S., Leong H.S., Lee D., Sellers R., Brullé-Soumaré L., Cairo S., Monteverde T., Volinia S., Smith D.D., Di Leva G., Galuppini F., Paliouras A.R., Zeng K., O’Keefe R, & Garofalo M. (2021). A KRAS-responsive long non-coding RNA controls microRNA processing. Nature Communications, 12, 2038.
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