AAV-SYN-NLS-SaCas9-NLS-3xHA-bGHpA-U6-BsaI-sgRNA
(Plasmid #218710)

• Purpose: AAV vector plasmid expressing Cas9 from Staphylococcus aureus (SaCas9) under the human synapsin (SYN) promoter and its sgRNA under the U6 promoter
• Depositing Lab: Bart Nieuwenhuis
• Publication: Adeno-associated viral vector-mediated delivery of SaCas9 in neurons (unpublished)

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Backbone

• Vector backbone: Addgene plasmid #61591
• Modifications to backbone: The CMV promoter has been substituted with the human synapsin (SYN) promoter between the AgeI and XbaI restriction enzyme digestion sites.
• Vector type: Mammalian Expression, AAV, CRISPR

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: hSaCas9
• Alt name: hSauCas9
• Species: Staphylococcus aureus
• Insert Size (bp): 3345
• GenBank ID: CCK74173.1
• Tags / Fusion Proteins:
  • NLS (N terminal on backbone)
  • NLS (C terminal on insert)
  • 3xHA (C terminal on insert)

Gene/Insert 2

• Gene/Insert name: Chimeric guide for SaCas9
• Species: Staphylococcus aureus
• Insert Size (bp): 101

Resource Information

• Supplemental Documents:
  • AAV-SYN-NLS-SaCas9-NLS-3xHA-bGHpA-U6-BsaI-sgRNA.gb

Terms and Licenses

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

Depositor Comments

This plasmid incorporates the synapsin promoter for neuron-specific and strong expression (DOI: 10.1038/s41434-020-0169-1 and 10.1038/s41434-022-00380-z). Further details on sgRNA cloning into this vector are available on the Addgene page of plasmid #61591.