AiP13044 - pAAV-AiE0779m_3xC2-minBG-SYFP2-WPRE3-BGHpA (Alias: CN3044)
(Plasmid #191706)

• Purpose: AI plasmid ALIAS: AiP13044. Optimized expression of SYFP2 (yellow fluorescent protein) in striatal direct pathway medium spiny neurons (D1-MSNs)
• Depositing Labs: The Allen Institute for Brain Science, Jonathan Ting
• Publication: Enhancer AAV toolbox for accessing and perturbing striatal cell types and circuits (unpublished)

Backbone

• Vector backbone: rAAV
• Vector type: AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SYFP2
• Species: Other
• Mutation: none

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)

Resource Information

• Supplemental Documents:
  • CN3044.gb

Terms and Licenses

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

Depositor Comments

Allen Institute plasmid ALIAS: AiP13044
This work supported by NIH BRAIN Initiative grant UF1MH128339

Information for AAV PHP.eB (Catalog # 191706-PHPeB)

Purpose

Ready-to-use AAV PHP.eB particles produced from AiP13044 - pAAV-AiE0779m_3xC2-minBG-SYFP2-WPRE3-BGHpA (Alias: CN3044) (#191706). In addition to the viral particles, you will also receive purified AiP13044 - pAAV-AiE0779m_3xC2-minBG-SYFP2-WPRE3-BGHpA (Alias: CN3044) plasmid DNA.

SYFP2-expression in striatal direct pathway medium spiny neurons (D1-MSNs). These AAV were produced with the PHPeB serotype, which permits efficient transduction of the central nervous system. These AAV preparations are suitable purity for injection into animals.

Delivery

• Volume: 100 µL
• Titer: ≥ 1×10¹³ vg/mL
• Pricing: $375 USD for preparation of 100 µL virus + $30 USD for plasmid.
• Storage: Store at -80℃. Thaw just before use and keep on ice.
• Shipment: Viral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.

Viral Production & Use

• Packaging Plasmids: encode adenoviral helper sequences and AAV rep gene, PHP.eB cap gene
  pUCmini-iCAP-PHP.eB (plasmid #103005)
• Buffer: PBS + 0.001% Poloxamer 188
• Serotype: PHPeB (plasmid #103005)
• Purification: Iodixanol gradient ultracentrifugation
• Reporter Gene: SYFP2

Biosafety

Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. Biosafety Guide

Terms and Licenses

• Academic/Nonprofit Terms:
  • Ancillary Agreement for Penn Vectors
  • Terms of Use for Viral Vectors
• Industry Terms:
  • Not Available to Industry

Viral Quality Control

Quality Control:

• Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). The specific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
• Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.

Visit our viral production page for more information.

Addgene Comments

Citation Information: When using the PHP.eB serotype in future publications, please acknowledge Viviana Gradinaru and cite Chan et al., Nat Neurosci, 20(8):1172-1179. Pubmed.