Pbabe puro mcherry egfp lc3

Manufactured by Addgene

Sourced in United States

PBabe-puro-mCherry-EGFP-LC3 is a plasmid construct that expresses mCherry-EGFP-LC3 fusion protein under the control of a puromycin resistance gene. This construct is designed for monitoring autophagy flux in cells.

Automatically generated - may contain errors

Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Egfp lc3, by Addgene (1 mentions) Plko 1 lentiviral vectors for shrnas, by Merck Group (1 mentions) Pbabe hygro, by Addgene (1 mentions) Plko 1 blast, by Addgene (1 mentions) Pspax2, by Addgene (1 mentions) Shc002, by Merck Group (1 mentions) Rab27a, by Merck Group (1 mentions) Pmd2 g, by Addgene (1 mentions)

Spelling variants of this product

PBABE-puro (113 citations) EGFP-LC3 (21 citations) MCherry-EGFP-LC3 (2 citations) PBABE-EGFP (2 citations)

2 protocols using pbabe puro mcherry egfp lc3

Plasmid Acquisition and Transfection

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The following plasmids were obtained from Addgene (Cambridge, MA USA) and directly used in the study: EGFP- microtubule-associated protein light chain 3 (EGFP-LC3) (Plasmid #11546), pBabe-puro-mCherry-EGFP-LC3 (plasmid #22418) and mouse kif5B (pKin1B, plasmid #31604). Kif5B were PCR amplified and subcloned into pEGFP-C1 vector. pcDNA3.1-full length htt23Q (fhtt23Q) and pcDNA3.1-fhtt145Q were originally obtained from HD Community BioRepository Collection. EGFP was engineered to the C-terminus of fhtt23Q (fhtt23Q-EGFP) and fhtt145Q (fhtt145Q-EGFP). Cells were transiently transfected with different plasmids using Lipofectamine 2000 (Invitrogen, Grand Island, NY, USA).

Erie C., Sacino M., Houle L., Lu M, & Wei J. (2015). Altered lysosomal positioning affects lysosomal functions in a cellular model of Huntington's disease. The European journal of neuroscience, 42(3), 1941-1951.

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Cloning and Lentiviral Constructs

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The following vectors are available or were obtained from Addgene: pBABE-puro mCherry-EGFP-LC3 (#22418), pBABE-hygro (#1765), psPAX2 (#12260), pMD2.G (Plasmid #12259), pLKO.1-blast (#26655), and pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro (#141383). pBABE-hygro mCherry-EGFP-LC3 was generated by subcloning mCherry-EGFP-LC3 from pBABE-puro mCherry-EGFP-LC3 into shared AgeI and SalI restriction sites in pBABE-hygro. pLKO.1 lentiviral vectors for shRNAs targeting SNAP29 (#1 TRCN0000083650; #2 TRCN0000231850), VAMP8 (#1 TRCN0000219911; #2 TRCN0000181123), Rab27a (#1 TRCN0000005295; #2 TRCN0000005296), and non-targeting control (SHC002) are available from Sigma-Aldrich. pLKO.1-blast shRab27a was generated by subcloning the blasticidin resistance gene from pLKO.1-blast into shared BamHI and KpnI restriction sites in pLKO.1 shRab27a (#TRCN0000005295).

Solvik T.A., Nguyen T.A., Tony Lin Y.H., Marsh T., Huang E.J., Wiita A.P., Debnath J, & Leidal A.M. (2022). Secretory autophagy maintains proteostasis upon lysosome inhibition. The Journal of Cell Biology, 221(6), e202110151.

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