PurposeHomology arms and linker-mEGFP sequence for internal insertion of CAGGS-mEGFP at the AAVS1 safe harbor location in human cells as a reporter for cytoplasmic mEGFP
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||91565||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector typeMammalian Expression, CRISPR ; Donor Template
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameAAVS1 Homology Arms with CAGGS driven mEGFP
SpeciesH. sapiens (human)
Insert Size (bp)5944
Entrez GeneAAVS1 (a.k.a. AAV)
- Promoter CAG
- Cloning method Restriction Enzyme
- 5′ cloning site unknown (unknown if destroyed)
- 3′ cloning site unknown (unknown if destroyed)
- 5′ sequencing primer unknown (Common Sequencing Primers)
Terms and Licenses
This plasmid has been used with locus-specific CRISPR/Cas9 to insert a CAGGS-mEGFP at the AAVS1 safe harbor locus in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. After insertion of the reporter using this donor template plasmid and CRISPR/Cas9 reagents, transfected cells may exhibit varying intensity levels of fluorescence, likely due to editing precision. To obtain cells of uniform intensity levels, see our protocol for fluorescence-assisted cell sorting and subcloning of transfected cells (https://www.allencell.org/instructional-videos-and-tutorials-for-cell-methods.html). Further, we recommend PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209). For more information on the entire plasmid collection, please see https://www.addgene.org/allen-institute-cell-science/ .
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:AICSDP-35:AAVS1-mEGFP was a gift from The Allen Institute for Cell Science (Addgene plasmid # 91565 ; http://n2t.net/addgene:91565 ; RRID:Addgene_91565)