Mucolipin1 pegfp c3

Manufactured by Addgene

Mucolipin1-pEGFP C3 is a plasmid construct that contains the gene encoding the mucolipin 1 protein fused to the enhanced green fluorescent protein (EGFP) tag. The plasmid is designed for the expression and visualization of the mucolipin 1 protein in mammalian cell lines.

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Lab products found in correlation

Plenti cmv mcs gfp sv puro, by Addgene (2 mentions) Pspax2, by Addgene (2 mentions) Pmd2 g, by Addgene (2 mentions) Prk5 ha gst ragc 75l, by Addgene (1 mentions)

Spelling variants of this product

PEGFP-C3 (3 citations)

2 protocols using mucolipin1 pegfp c3

Generating EGFP-MCOLN1 Lentiviral Construct

Cited 1 time

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To obtain EGFP-MCOLN1 construct for lentiviral transduction, human MCOLN1 was amplified by PCR using the following oligonucleotides with overhangs (underlined): 5′-GACACCGACTCTAGAATGGTGAGCAAGGGCGAGGAGC-3′ (forward) and 5′-AACTAGTCCGGATCCTCAATTCACCAGCAGCGAATGC-3′ (reverse) from the mucolipin1-pEGFP C3 (plasmid #62960; Addgene) construct and subcloned into XbaI and BamHI restriction sites of the pLenti-CMV-MCS-GFP-SV-puro (plasmid #73582; Addgene) vector using the sequence- and ligation-independent cloning (SLIC) method as described elsewhere (Jeong et al, 2012 (link)). The mucolipin1-pEGFP C3 (plasmid #62960; Addgene) construct was a gift from Paul Luzio (Pryor et al, 2006 (link)). pRK5-HA GST RagC WT (plasmid #19304; Addgene) and pRK5-HA GST RagC 75L (plasmid #19305; Addgene) were a gift from David Sabatini (Sancak et al, 2008 (link)). psPAX2 (plasmid #12260; Addgene) and pMD2.G (plasmid #12259; Addgene) lentiviral packaging plasmids were a gift from Didier Trono.

Wróbel M., Cendrowski J., Szymańska E., Grębowicz-Maciukiewicz M., Budick-Harmelin N., Macias M., Szybińska A., Mazur M., Kolmus K., Goryca K., Dąbrowska M., Paziewska A., Mikula M, & Miączyńska M. (2022). ESCRT-I fuels lysosomal degradation to restrict TFEB/TFE3 signaling via the Rag-mTORC1 pathway. Life Science Alliance, 5(7), e202101239.

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Generating EGFP-MCOLN1 lentiviral construct

Check if the same lab product or an alternative is used in the 5 most similar protocols

To obtain EGFP-MCOLN1 construct for lentiviral transduction, human MCOLN1 was amplified by PCR with the following oligonucleotides with overhangs (underlined): 5'-GACACCGACTCTAGAATGGTGAGCAAGGGCGAGGAGC-3' (forward) and 5'-AACTAGTCCGGATCCTCAATTCACCAGCAGCGAATGC-3' (reverse) from Mucolipin1-pEGFP C3 (Addgene plasmid #62960) construct and subcloned into XbaI and BamHI restriction sites of pLenti-CMV-MCS-GFP-SV-puro (Addgene plasmid #73582) vector using sequence-and ligation-independent cloning (SLIC) method as described elsewhere (Jeong et al, 2012) . Mucolipin1-pEGFP C3 (Addgene plasmid#62960) construct was a gift from Paul Luzio (Pryor et al, 2006) . psPAX2 (Addgene plasmid # 12260) and pMD2.G (Addgene plasmid # 12259) lentiviral packaging plasmids were a gift from Didier Trono.

Szymańska E., Szymańska E., Budick-Harmelin N., Kolmus K., Goryca K., Dąbrowska M., Paziewska A., Mikula M., Cendrowski J., & Miączyńska M. (2021). ESCRT-I controls lysosomal membrane protein homeostasis and restricts MCOLN1-dependent TFEB/TFE3 signaling.

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