Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein
Type
Blog Post
...pitfalls: they require construction of plasmid vectors and/or rely on cumbersome in vitro synthesis of... have low knock-in efficiency. Together, these factors prevent stem cell researchers from generating clonal...purchased and there is no need to construct plasmid vectors or synthesize/purify IVT sgRNA in the lab.
Figure...experiments. Notably, we could tag >60 transcription factors in a short (3-week) time window; this was possible...csRNP experiments, it can be ordered in bulk and stored in small aliquots at -80 oC.
Assemble csRNP: Anneal...success of knock-in experiments depends on many factors. Some of these include:
Most importantly, the...