Pig-GESS004
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Pig-GESS004

Gene editing activity testing

Format - cell-based transfection

Assay - T7 endonuclease I/Cel-II/Surveyor
  • Quantity
Pig-GESS004
Product Details
NovoHelix offers a gene editing service to help clients test their CRISPR tools including guide RNAs, high-performance mutant Cas proteins and base editors in plasmid DNA or RNP formats.  A representative cell line will be transfected in triplicate and results will be generated by the mismatch-nucleases T7 Endo I or Cel-II as adopted from a protocol originally developed by Keith Joung's lab. While our cell-based assay is the gold standard for guide RNA activity, guide RNAs can be tested in an alternative in vitro cutting assay should the client prefer.  However, caution must be used in interpreting these in vitro results, as NovoHelix has broadly found that Cas9 RNP in vitro cutting of PCR amplicons as a surrogate assay for gRNA activity vastly overrepresents Cas RNP in vivo activity levels both in cellular and microinjection contexts. Hence in vitro mismatch nuclease assays often do not correlate to gene editing in vivo outcomes. In short, the cell-based gRNA testing is strongly suggested before proceeding to animal model generation.