Prof: Your CRISPR editing experiment worked?
Student: CRISPR won’t EDIT, it only CUTS✂️
P: Who edits then?
S: Cell does. But it is unpredictable
P: How can you find out?
S: Using CRISPR-KRISPR
P: What?
S: Yeah, CRISPR-KRISPR, a new method. See paper https://genomebiology.biomedcentral.com/articles/10.1186/s13059-022-02779-8
36s
CRISPRGuru @CRISPRGuru
🧵The CRISPR-KRISPR method was a collaborative work from
@Masato_Ohtsuka
lab. His talented team of bioinformaticians came up with a strategy to simplify Circle-Seq, a popular approach for detecting off target editing events
nature.com
CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR–Cas9 nuclease off-targets
https://www.nature.com/articles/nmeth.4278
We named our method as CRISPR-KRISPR (CRISPR-Knock-ins and Random Inserts Searching PRotocol)
The method uses very less DNA and it can do much more than just finding out off target cleavages in the genome.
We did not find off target cleavages in genetically engineered mouse (GEM) models. It is already well established that off-target cleavages are not common in GEM mice. It is nicely documented in this paper. https://www.nature.com/articles/nmeth.3408