AICSDP-167:CDH5-mEGFP
(Plasmid #217425)

• Purpose: Homology arms and mEGFP sequence for C-terminus tagging of human cadherin 5
• Depositing Lab: Allen Institute for Cell Science
• Publication: Allen Institute for Cell Science Plasmids (unpublished)

Backbone

• Vector backbone: pAAV
• 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: CDH5 Homology Arms with mEGFP
• Alt name: cadherin 5
• Species: H. sapiens (human)
• Insert Size (bp): 2714
• Mutation: homology arms contain point mutations to disrupt crRNA binding sites used, and WTC-11 specific homozygous SNP(s)
• Entrez Gene: CDH5 (a.k.a. 7B4, CD144)
• Tag / Fusion Protein:
  • mEGFP (N terminal on insert)

Cloning Information

• Cloning method: Unknown

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

Depositor Comments

This plasmid has been used with locus-specific CRISPR/Cas9 to add a mEGFP tag to the C-terminus of human CDH5 in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. After initial HDR, we recommend subcloning of transfected cells and PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209) After identifying precisely edited clones, differentiate WTC human induced pluripotent stem cells to endothelial cells to observe cells expressing mEGFP tagged CDH5.