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pAAV-Syn-ChR2(H134R)-GFP
(Plasmid #58880)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 58880 Standard format: Plasmid sent in bacteria as agar stab 1 $65
AAV8 58880-AAV8 Virus (100 µL at titer ≥ 7×10¹² vg/mL) and Plasmid. More Information
$380
AAV Retrograde 58880-AAVrg Virus (100 µL at titer ≥ 7×10¹² vg/mL) and Plasmid. More Information
$380

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    AAV
  • Backbone size w/o insert (bp) 4698
  • Total vector size (bp) 6360
  • Vector type
    Mammalian Expression, AAV

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Growth instructions
    Stbl3 at 30C (use carbenicillin if using Stbl3) OR DH5a at 37C
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    ChR2(H134R)-GFP
  • Alt name
    channelrhodopsin-2 H134R mutant
  • Species
    Chlamydomonas reinhardtii
  • Insert Size (bp)
    1662
  • Mutation
    H134R
  • GenBank ID
    EU714030.1
  • Promoter Syn
  • Tag / Fusion Protein
    • GFP (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site EcoRI (not destroyed)
  • 5′ sequencing primer CTGCGTATGAGTGCAAG
  • 3′ sequencing primer cagcgtatccacatagcg
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Plasmid is completely sequenced by the depositing lab. Multiple digestions were done to verify the vector structure. The construct and the virus were both tested in vitro.

Information for AAV8 (Catalog # 58880-AAV8) ( Back to top )

Purpose

Ready-to-use AAV8 particles produced from pAAV-Syn-ChR2(H134R)-GFP (#58880). In addition to the viral particles, you will also receive purified pAAV-Syn-ChR2(H134R)-GFP plasmid DNA.

These AAV preparations are suitable purity for injection into animals.

Delivery

  • Volume 100 µL
  • Titer ≥ 7×10¹² vg/mL
  • Pricing $350 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, AAV8 cap gene
  • Buffer PBS + 0.001% Pluronic F-68
  • Serotype AAV8
  • Purification Iodixanol gradient ultracentrifugation
  • Reporter Gene GFP

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

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.

Information for AAV Retrograde (Catalog # 58880-AAVrg) ( Back to top )

Purpose

Ready-to-use AAV Retrograde particles produced from pAAV-Syn-ChR2(H134R)-GFP (#58880). In addition to the viral particles, you will also receive purified pAAV-Syn-ChR2(H134R)-GFP plasmid DNA.

Humanized channelrhodopsin H134R mutant fused to GFP, under the control of the Synapsin promoter. These AAV were produced with a retrograde serotype, which permits retrograde access to projection neurons. These AAV preparations are suitable purity for injection into animals.

Delivery

  • Volume 100 µL
  • Titer ≥ 7×10¹² vg/mL
  • Pricing $350 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, AAV retrograde cap gene from rAAV2-retro helper (plasmid #81070)
  • Buffer PBS + 0.001% Pluronic F-68 + 200 mM NaCl
  • Serotype AAV retrograde (AAVrg)
  • Purification Iodixanol gradient ultracentrifugation
  • Reporter Gene GFP

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

Resource Information

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

Retrograde functionality is dependent on high viral titers. Addgene recommends not diluting your AAV preps prior to use.
How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pAAV-Syn-ChR2(H134R)-GFP was a gift from Edward Boyden (Addgene plasmid # 58880)

    For viral preps, please replace (Addgene plasmid # 58880) in the above sentence with: (Addgene viral prep # 58880-AAV8) or (Addgene viral prep # 58880-AAVrg)

  • For your References section:

    Millisecond-timescale, genetically targeted optical control of neural activity. Boyden ES, Zhang F, Bamberg E, Nagel G, Deisseroth K. Nat Neurosci. 2005 Sep . 8(9):1263-8. 10.1038/nn1525 PubMed 16116447