Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein
Type
Blog Post
...IP and ChIP assays.
Tag 96 genes at a time using Custom Synthetic cr:tracrRNA/Cas9 Ribonucleoprotein (csRNP...first step of optimization, we started off by using custom synthetic two-part cr/tracrRNAs for CRISPR experiments...tagging, pick a crRNA that cuts 8-15 bp downstream of Stop codon; for N-terminus tagging, a crRNA cutting 8...csRNP experiments, it can be ordered in bulk and stored in small aliquots at -80 oC.
Assemble csRNP: Anneal...near the insertion site (within ~8-15 bp from the stop codon for C-terminus tagging)
GC content of the ... hands, delivering Cas9 protein in complex with custom synthetic cr:tracrRNAs (csRNP) is the most efficient...