Lzrs ires gfp

Manufactured by Addgene

The LZRS-IRES-GFP is a retroviral vector that allows for the expression of a gene of interest and Green Fluorescent Protein (GFP) from a single transcript. It contains an Internal Ribosome Entry Site (IRES) sequence that enables the translation of two open reading frames from the same mRNA. This vector can be used for the stable transduction of mammalian cells and the identification of successfully transduced cells through the expression of GFP.

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Lab products found in correlation

Tet plko puro, by Addgene (1 mentions) Lenticas9 blast, by Addgene (1 mentions) Pl crispr, by Addgene (1 mentions) Quikchange 2 site directed mutagenesis kit, by Agilent Technologies (1 mentions)

2 protocols using lzrs ires gfp

Generating Genetically Modified Cell Lines

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The MYD88 coding sequence was subcloned into LZRS-IRES-GFP (Addgene plasmid #21961). Subsequently, the QuikChange II Site-Directed Mutagenesis Kit (Agilent, Santa Clara, California, USA) was utilized to generate the MYD88 mutant constructs according to the manufacturer’s instructions. Mutants were confirmed by Sanger sequencing. To generate doxycycline-inducible MYD88 knockdown cell lines, we inserted an shRNA targeting MYD88 (GCAGAGCAAGGAATGTGACTT or GACCCAATGTACCAGTATT) into Tet-pLKO-puro (Addgene plasmid #21915). To generate MYD88 knockout cells, we inserted a single guide RNA targeting MYD88 (CTGCTCTCAACATGCGAGTG or CTCGAGCAGTCGGCCTACAG) into pL-CRISPR.EFS.GFP (Addgene plasmid #57818). For generation of stable Cas9 expressing cell lines, we used lentiCas9-Blast (Addgene plasmid #52962). MSCV-CA-IKK2-IRES-GFP was kindly provided by Dr J. Schuringa (University of Groningen, Groningen, The Netherlands).

Minderman M., Lantermans H., van der Zwaan C., Hoogendijk A.J., van den Biggelaar M., Kersten M.J., Spaargaren M, & Pals S.T. (2023). The oncogenic human B-cell lymphoma MYD88 L265P mutation genocopies activation by phosphorylation at the Toll/interleukin-1 receptor (TIR) domain. Blood Cancer Journal, 13(1), 125.

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Stable Transfection of NEDD9 in Colon Cancer Cells

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Plasmid LZRS/IresGFP, LZRS/IresGFP-hNEDD9, pLKO, pLKO/shC, and pLKO/shD were from Addgene (Cambridge, MA). DLD1 and SW480 cells were transfected with 4 μg plasmid using Lipofectamine. Cells that stably expressing or knockdown NEDD9 were selected using 10 μg/mL puromycin for 2 weeks followed by immunoblotting analysis to verify NEDD9 expression.

Dai J., Van Wie P.G., Fai L.Y., Kim D., Wang L., Poyil P., Luo J, & Zhang Z. (2016). Downregulation of NEDD9 by apigenin suppresses migration, invasion, and metastasis of colorectal cancer cells. Toxicology and applied pharmacology, 311, 106-112.

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