pAAV-CMVie-NcanGFP
(Plasmid #235983)

• Purpose: Expresses the hyaluronan-binding domain of Neurocan fused to eGFP under the CMV promoter. It contains a sequence for secretory pathway, so the probe is exported to the extracellular space
• Depositing Lab: Federico Soria
• Publication: Viral-mediated fluorescent labeling of hyaluronan reveals extracellular matrix dynamics in the mouse brain in vivo (unpublished)

Backbone

• Vector backbone: pAAV-CMVie
• Backbone size w/o insert (bp): 3721
• Total vector size (bp): 7014
• Modifications to backbone: WPRE
• Vector type: AAV

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: Neurocan-GFP
• Alt name: NcanGFP
• Species: M. musculus (mouse)
• Insert Size (bp): 3293
• Promoter: CMVie
• Tag / Fusion Protein:
  • GFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AflII (unknown if destroyed)
• 3′ cloning site: MIuI (unknown if destroyed)
• 5′ sequencing primer: Unknown

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Kelly Smith, University of Melbourne ()

Terms and Licenses

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

Depositor Comments

Dr. Smith cloned the gene and used it in zebrafish (De Angelis et al., 2017, Dev Cell). She sent us the plasmid back in 2018. We cloned it into an AAV vector, and added the WPRE sequence. We have also validated its use in mouse and rat, and checked that it labels hyaluronan without affecting its properties and metabolism (Fernandez-Ballester et al., 2025, bioRxiv).

Please visit https://doi.org/10.1101/2025.01.31.635882 for bioRxiv preprint.