plko-shmfn2-mito-GFP-ER tethering
(Plasmid #160509)

• Purpose: Rescue mfn2 knockdown by tethering mitochondria and ER with GFP
• Depositing Lab: Qing Deng
• Publication: Zhou et al J Cell Sci. 2020 Sep 4;133(17). pii: jcs.248880. doi: 10.1242/jcs.248880.

Backbone

• Vector backbone: pLKO.1 lentiviral backbone
• Backbone manufacturer: sigma MISSION
• Backbone size w/o insert (bp): 8500
• Modifications to backbone: GFP was removed from the backbone (MFN2-sh2: TRCN0000082687) and the tether rescue construct was inserted.
• Vector type: Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: mito-GFP-ER tethering
• Species: H. sapiens (human)
• Insert Size (bp): 1274
• Promoter: CMV

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: CTCCCGGTCGCCACC ATGGCAATCCAGTTGCGTTCG
• 3′ sequencing primer: TCGAGGTCGAGAATT TTA AGATACATTGATGAGTTTGG

Resource Information

• Supplemental Documents:
  • plko-shmfn2-ER-m-GFP tether.gb
• A portion of this plasmid was derived from a plasmid made by: pLKO.1 lentiviral constructs with shRNA (TRCN0000082687) was from MILLIPORE SIGMA.
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The mitochondria-GFP-ER tether construct was published. The GFP containing a mitochondria localization signal (ATGGCAATCCAGTTGCGTTCGCTCTTCCCCTTGGCATTGCCCGGAATGCTGGCCCTCCTTGGCTGGTGGTGGTTTTTCTCTCGTAAAAAA) and ER localization signal (ATGGTTTATATTGGCATCGCTATTTTTTTGTTTTTGGTGGGCCTGTTTATGAAA) at it N- and C- terminal respectively.
1. Kornmann, B. et al. An ER-mitochondria tethering complex revealed by a synthetic biology screen. Science (80-. ). 325, 477–481 (2009).