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PurposeAAV expression of humanized ChR2(H134R) fused to EYFP driven by CaMKIIa promoter for optogenetic activation
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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 26969 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
AAV1 | 26969-AAV1 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid. | $405 | ||
AAV2 | 26969-AAV2 | Virus (100 µL at titer ≥ 3×10¹² vg/mL) and Plasmid. | $405 | ||
AAV5 | 26969-AAV5 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid. | $405 | ||
AAV9 | 26969-AAV9 | Virus (100 µL at titer ≥ 1×10¹³ vg/mL) and Plasmid. | $405 |
Backbone
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Vector backbonepAAV
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5390
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Vector typeMammalian Expression, AAV
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)Stbl3
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Growth instructionsStbl3 (rec A-) cells to avoid recombinations with the LTRs
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namehChR2(H134R)
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Alt nameChR2
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SpeciesH. sapiens (human)
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Insert Size (bp)1662
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MutationHistidine 134 is mutated to Arginine
- Promoter CaMKIIa
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Tag
/ Fusion Protein
- EYFP (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI, SalI, KpnI (not destroyed)
- 3′ cloning site EcoRI, HinDIII (not destroyed)
- 5′ sequencing primer ctcagaagccccaagctcgtc
- 3′ sequencing primer GCAATAGCATGATACAAAGG (Common Sequencing Primers)
Resource Information
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Articles Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
This plasmid contains the mouse calcium/calmodulin-dependent kinase II alpha subunit promoter.
For additional information please visit - http://www.optogenetics.org
Information for AAV1 (Catalog # 26969-AAV1) ( Back to top)
Purpose
Ready-to-use AAV1 particles produced from pAAV-CaMKIIa-hChR2(H134R)-EYFP (#26969). In addition to the viral particles, you will also receive purified pAAV-CaMKIIa-hChR2(H134R)-EYFP plasmid DNA.
CaMKIIa-driven, humanized channelrhodopsin H134R mutant fused to EYFP for optogenetic activation. 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, AAV1 cap gene
- Buffer PBS + 0.001% Poloxamer 188
- Serotype AAV1
- Purification Iodixanol gradient ultracentrifugation
- Reporter Gene EYFP
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
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Viral 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
Data submitted about 26969-AAV1 by requesting scientist(s):
Information for AAV2 (Catalog # 26969-AAV2) ( Back to top)
Purpose
Ready-to-use AAV2 particles produced from pAAV-CaMKIIa-hChR2(H134R)-EYFP (#26969). In addition to the viral particles, you will also receive purified pAAV-CaMKIIa-hChR2(H134R)-EYFP plasmid DNA.
CaMKIIa-driven, humanized channelrhodopsin H134R mutant fused to EYFP for optogenetic activation. These AAV preparations are suitable purity for injection into animals.Delivery
- Volume 100 µL
- Titer ≥ 3×10¹² 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, AAV2 cap gene
- Buffer PBS + 0.001% Poloxamer 188
- Serotype AAV2
- Purification Iodixanol gradient ultracentrifugation
- Reporter Gene EYFP
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
-
Industry Terms
- Not Available to Industry
Viral 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 AAV5 (Catalog # 26969-AAV5) ( Back to top)
Purpose
Ready-to-use AAV5 particles produced from pAAV-CaMKIIa-hChR2(H134R)-EYFP (#26969). In addition to the viral particles, you will also receive purified pAAV-CaMKIIa-hChR2(H134R)-EYFP plasmid DNA.
CaMKIIa-driven, humanized channelrhodopsin H134R mutant fused to EYFP for optogenetic activation. 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, AAV5 cap gene
- Buffer PBS + 0.001% Poloxamer 188
- Serotype AAV5
- Purification Iodixanol gradient ultracentrifugation
- Reporter Gene EYFP
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
-
Industry Terms
- Not Available to Industry
Viral 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 AAV9 (Catalog # 26969-AAV9) ( Back to top)
Purpose
Ready-to-use AAV9 particles produced from pAAV-CaMKIIa-hChR2(H134R)-EYFP (#26969). In addition to the viral particles, you will also receive purified pAAV-CaMKIIa-hChR2(H134R)-EYFP plasmid DNA.
CaMKIIa-driven, humanized channelrhodopsin H134R mutant fused to EYFP for optogenetic activation. 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, AAV9 cap gene
- Buffer PBS + 0.001% Poloxamer 188
- Serotype AAV9
- Purification Iodixanol gradient ultracentrifugation
- Reporter Gene EYFP
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
-
Industry Terms
- Not Available to Industry
Viral 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
Data submitted about 26969-AAV9 by requesting scientist(s):
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.
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For your Materials & Methods section:
pAAV-CaMKIIa-hChR2(H134R)-EYFP was a gift from Karl Deisseroth (Addgene plasmid # 26969 ; http://n2t.net/addgene:26969 ; RRID:Addgene_26969) For viral preps, please replace (Addgene plasmid # 26969) in the above sentence with: (Addgene viral prep # 26969-AAV1), (Addgene viral prep # 26969-AAV2), (Addgene viral prep # 26969-AAV5), or (Addgene viral prep # 26969-AAV9) -
For your References section:
Global and local fMRI signals driven by neurons defined optogenetically by type and wiring. Lee JH, Durand R, Gradinaru V, Zhang F, Goshen I, Kim DS, Fenno LE, Ramakrishnan C, Deisseroth K. Nature. 2010 Jun 10. 465(7299):788-92. 10.1038/nature09108 PubMed 20473285