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Addgene/pAAV-Ef1a-DIO hChR2(E123T/T159C)-EYFP/1unit/35509-AAV9

Ordering

ItemCatalog #DescriptionQuantityPrice (USD)
Plasmid35509Standard format: Plasmid sent in bacteria as agar stab1$75
Add to Cart
AAV535509-AAV5Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information
$380
Add to Cart
AAV935509-AAV9Virus (100 µL at titer ≥ 1×10¹³ vg/mL)and Plasmid.More Information
$380
Add to Cart

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pAAV
    (Search Vector Database)
  • Backbone manufacturer
    Stratagene
  • Backbone sizew/o insert(bp)5605
  • Modifications to backbone
    Addition of an Ef1a promoter, lox sites and WPRE
  • Vector type
    Mammalian Expression, AAV

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    hChR2
  • Species
    Chlamydomonas reinhardtii
  • Insert Size (bp)
    1661
  • Mutation
    E123T and T159C
  • PromoterEf1a
  • Tag/ Fusion Protein
    • EYFP (C terminal on insert)

Cloning Information

  • Cloning methodRestriction Enzyme
  • 5′ cloning siteAscI(not destroyed)
  • 3′ cloning siteNheI(not destroyed)
  • 5′ sequencing primerCACCCACACAAAGGAAAAGGGCC
  • 3′ sequencing primerGCAATAGCATGATACAAAGG
    • (Common Sequencing Primers)

Resource Information

  • Terms and Licenses
    • UBMTA
    • Ancillary Agreement for Plasmids Containing FP Materials
    • genOway Notice of RIghts
  • Industry Terms
    • Not Available to Industry
  • Articles Citing this Plasmid
    • 6 References

Information for AAV5 (Catalog # 35509-AAV5)(Back to top)

Purpose

Ready-to-use AAV5 particles produced from pAAV-Ef1a-DIO hChR2(E123T/T159C)-EYFP (#35509). In addition to the viral particles, you will also receive purified pAAV-Ef1a-DIO hChR2(E123T/T159C)-EYFP plasmid DNA.

EF1a-driven, Cre-dependent, humanized channelrhodopsin E123T/T159C mutant fused to EYFP for optogenetic activation.These AAV preparations are suitable purity for injection into animals.

Delivery

  • Volume100 µL
  • Titer≥ 1×10¹³ vg/mL
  • Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
  • StorageStore at -80℃. Thaw just before use and keep on ice.
  • ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.

Viral Production & Use

  • Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV5 cap gene
  • BufferPBS + 0.001% Pluronic F-68
  • SerotypeAAV5
  • PurificationIodixanol gradient ultracentrifugation
  • Reporter GeneEYFP (Cre-dependent)

Biosafety

Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide

Resource Information

  • Terms and Licenses
    • 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). Thespecific 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 moreinformation.

Addgene Comments

Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.

Information for AAV9 (Catalog # 35509-AAV9)(Back to top)

Purpose

Ready-to-use AAV9 particles produced from pAAV-Ef1a-DIO hChR2(E123T/T159C)-EYFP (#35509). In addition to the viral particles, you will also receive purified pAAV-Ef1a-DIO hChR2(E123T/T159C)-EYFP plasmid DNA.

EF1a-driven, Cre-dependent, humanized channelrhodopsin E123T/T159C mutant fused to EYFP for optogenetic activation.These AAV preparations are suitable purity for injection into animals.

Delivery

  • Volume100 µL
  • Titer≥ 1×10¹³ vg/mL
  • Pricing$350 USD for preparation of 100 µL virus + $30 USD for plasmid.
  • StorageStore at -80℃. Thaw just before use and keep on ice.
  • ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.

Viral Production & Use

  • Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV9 cap gene
  • BufferPBS + 0.001% Pluronic F-68
  • SerotypeAAV9
  • PurificationIodixanol gradient ultracentrifugation
  • Reporter GeneEYFP (Cre-dependent)

Biosafety

Requestor is responsible for compliance withtheir institution"s biosafety regulations.Lentivirus is generally considered BSL-2. AAV isgenerally considered BSL-1, but may requireBSL-2 handling depending on the insert.Biosafety Guide

Resource Information

  • Terms and Licenses
    • 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). Thespecific 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 moreinformation.

Addgene Comments

Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.

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