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Pgfp c shlenti plasmid

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The PGFP-C-shLenti plasmid is a laboratory tool designed for gene knockdown experiments. It contains a green fluorescent protein (GFP) reporter and a short hairpin RNA (shRNA) expression cassette. This plasmid can be used to generate lentiviral particles for the delivery and expression of shRNA in target cells, enabling researchers to study the effects of gene silencing.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Sicontrol, by Horizon Discovery (1 mentions) Interferin transfection reagent, by Polyplus Transfection (1 mentions) M165 fc fluorescent microscope, by Leica (1 mentions)

Close spelling variants of this product

PGFP-C-shLenti

4 protocols using pgfp c shlenti plasmid

1

Immortalized Fibroblast Generation

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The pBABE-neo-hTERT plasmid (#1774, Addgene) was used to generate immortalized fibroblasts. NF47 and CAF47 cells were infected with retroviruses and passaged. hTERT levels were validated using RT-PCR. The pGFP-C-shLenti plasmid containing anti-TINAGL1 (TL307228, Origene) or scrambled negative control sequences (TR30023, OriGene) were used to generate stable cell lines. Immortalized CAFs were infected with lentivirus, and GFP-expressing cells were selected. TINAGL1 levels were validated by RT-PCR.
Lee D., Ham I.H., Oh H.J., Lee D.M., Yoon J.H., Son S.Y., Kim T.M., Kim J.Y., Han S.U, & Hur H. (2024). Tubulointerstitial nephritis antigen-like 1 from cancer-associated fibroblasts contribute to the progression of diffuse-type gastric cancers through the interaction with integrin β1. Journal of Translational Medicine, 22, 154.
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2

Efficient Gene Knockdown Techniques

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For stable knockdown (shRNA) of IDO1, ALC cells were transfected with 1μg of pGFP-C-shLenti plasmid (Origene) expressing shIDO1 (NM_002164) or scrambled control using lipofectamine 2000 (Thermo Fisher) transfection reagent (see Supplemental Method). After 24h, transfection medium Opti-MEM was exchanged to RPMI1640 containing 5μg/ml of G418. GFP as a reporter was used to evaluate target gene knocked down efficiency. For siRNA, cell lines A were transfected with 1nM of HIF1α SMARTpool® siRNA (NM_005566) or siCONTROL®(Dharmacon) using INTERFERin™ transfection reagent (Polyplus) as previously described (26 (link)).
Nguyen D.J., Theodoropoulos G., Li Y.Y., Wu C., sha W., Feun L.G., Lampidis T.J., Savaraj N, & Wangpaichitr M. (2019). Targeting the kynurenine pathway for the treatment of cisplatin resistant lung cancer. Molecular cancer research : MCR, 18(1), 105-117.
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3

Lentiviral-mediated SLC2A1 knockdown

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HCT-116 cells were infected with lentivirus containing either SLC2A1-specific short hairpin RNA or scrambled short hairpin RNA in pGFP-C-shLenti plasmid (OriGene). Infected cells were then plated into 96-well microdilution plates to measure cell viability over 6 days by MTT assay as previously described.
Lopez-Serra P., Marcilla M., Villanueva A., Ramos-Fernandez A., Palau A., Leal L., Wahi J.E., Setien-Baranda F., Szczesna K., Moutinho C., Martinez-Cardus A., Heyn H., Sandoval J., Puertas S., Vidal A., Sanjuan X., Martinez-Balibrea E., Viñals F., Perales J.C., Bramsem J.B., Ørntoft T.F., Andersen C.L., Tabernero J., McDermott U., Boxer M.B., Heiden M.G., Albar J.P, & Esteller M. (2014). A DERL3-associated defect in the degradation of SLC2A1 mediates the Warburg effect. Nature Communications, 5, 3608.
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4

Tumor Induction and Visualization on Chick CAM

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ISK, and HEC1A cells were labelled with an EGFP-expressing lentivirus donated by Dr Sokratis Theocharatos (University of Liverpool, UK). RL95-2 cells were labelled with a lentivirus expressing tGFP (pGFP-C-shLenti plasmid) (Origene, MD, USA), and selected using puromycin antibiotic selection. ISK and HEC1A viral transfection and egg preparation were performed according to Carter et al. 2012 [20] . Cells were added to the CAM at embryonic day 7 (E7) [21] the resulting tumours were imaged at E14, using a standard Leica M165-FC fluorescent microscope, in situ and after excision.
(Fig. 5C) the resulting tumours were fixed in 10% (v/v) NBF and embedded in paraffin wax.
Parkes C., Kamal A., Valentijn A.J., Alnafakh R., Gross S.R., Barraclough R., Moss D., Kirwan J, & Hapangama D.K. (2018). Assessing Estrogen-Induced Proliferative Response in an Endometrial Cancer Cell Line Using a Universally Applicable Methodological Guide. International journal of gynecological cancer : official journal of the International Gynecological Cancer Society, 28(1).
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