SAM-based Genome Editing to Obtain Isogenic Cell Lines Overexpressing Human CD5 Protein

In vitro and in vivo experiments often require construction of convenient cell instruments to reliably assess the specificity of the molecules or therapeutic approaches being tested against the protein target of interest. Using model isogenic cell lines that differ only in the expression of the target protein represents an ideal solution to this problem. Cloning and efficient delivery of genetic cassettes encoding such proteins, particularly the large ones, is typically challenging, much as the knock-out of the respective genes. To tackle this issue, we adapted a CRISPR/ Cas9-based SAM (Synergistic Activation Mediator) platform, and successfully established four model isogenic cell line pairs (U343, HeLa, HT-1080 и HEp-2) overexpressing human CD5.