Screen It™ CRISPR Cas9 Cleavage Detection Kit

Product Description

abm’s Screen It™ CRISPR Cas9 Cleavage Detection Kit is a robust and precise RNA-dependent assay used to identify successful insertions and/or deletions (indels) of genomic DNA following a CRISPR-Cas9 experiment. CRISPR-Cas9 can introduce indels on either one or both copies of a given mammalian gene. abm’s Screen It™ CRISPR Cas9 Cleavage Detection Kit can easily identify if a given clone is monoallelic (has mutations in one copy), biallelic (both copies mutated) or is unchanged wild-type which is considered a major screening advantage over the standard T7E1 Surveyor Assay.

This kit uses PCR to first amplify the sgRNA target site from CRISPR edited cells; this amplicon is subsequently cleaved by a ribonucleoprotein (RNP) complex which generates a unique pattern that can be easily resolved on agarose gel and interpreted by the user. Using this kit after gene editing saves time and energy from the arduous screening process and provides clear genotyping results.

Supporting Data

Figure 1. Screen It CRISPR Cas9 Cleavage Detection Kit Controls showing the distinct patterns after spCas9 cleavage. Wild-type Control (1) shows 2 bands - 194 bp + 209 bp, Monoallelic Control (2) shows 3 bands - 503 bp + 294 bp + 209 bp and Biallelic Control (3) shows 1 band - 503 bp.

Figure 2. RNP complex introduces a double-stranded break on Wild-type sample (blue) but not on CRISPR-edited samples (Red). Subsequent cleavage will show one cleaved band for Wild-type (2 fragments), one uncleaved and one cleaved band for Monoallelic editing (3 fragments) and one uncleaved band for Biallelic editing (1 fragment).