AICSDP-078:G3BP1-mEGFP
(Plasmid
#193920)
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PurposeHomology arms and mEGFP-SGLRS for N-terminus tagging of human G3BP stress granule assembly factor 1
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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 | 193920 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 * |
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Backbone
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Vector backbonepUC57
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Vector typeCRISPR ; Donor Template
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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameG3BP1 Homology Arms with mEGFP-SGLRS
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Alt nameG3BP stress granule assembly factor 1
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SpeciesH. sapiens (human)
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Insert Size (bp)2729
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Mutationhomology arms contain point mutations to disrupt crRNA binding sites used, and WTC-11 specific homozygous SNP(s)
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Entrez GeneG3BP1 (a.k.a. G3BP, HDH-VIII)
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Tag
/ Fusion Protein
- mEGFP-linker (N terminal on insert)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
This plasmid has been used with locus-specific CRISPR/Cas9 to add a mEGFP tag to the N-terminus of human G3BP1 in WTC-11 human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: SGLRS. After protein tagging 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.
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For your Materials & Methods section:
AICSDP-078:G3BP1-mEGFP was a gift from Allen Institute for Cell Science (Addgene plasmid # 193920 ; http://n2t.net/addgene:193920 ; RRID:Addgene_193920)