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Addgene

AICSDP-078:G3BP1-mEGFP
(Plasmid #193920)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 193920 Standard format: Plasmid sent in bacteria as agar stab 1 $85 *

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Backbone

  • Vector backbone
    pUC57
  • Vector type
    CRISPR ; Donor Template

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    G3BP1 Homology Arms with mEGFP-SGLRS
  • Alt name
    G3BP stress granule assembly factor 1
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    2729
  • Mutation
    homology arms contain point mutations to disrupt crRNA binding sites used, and WTC-11 specific homozygous SNP(s)
  • Entrez Gene
    G3BP1 (a.k.a. G3BP, HDH-VIII)
  • Tag / Fusion Protein
    • mEGFP-linker (N terminal on insert)

Terms and Licenses

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/ .

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:

    AICSDP-078:G3BP1-mEGFP was a gift from Allen Institute for Cell Science (Addgene plasmid # 193920 ; http://n2t.net/addgene:193920 ; RRID:Addgene_193920)