Application

CRISPR/Cas9 can be used to introduce, or “knock in”, new DNA sequences. Common modifications include the introduction of a SNP, small tag, loxP or larger cassette such as fluorescent protein. These modifications are made through the introduction of targeted DSB which enhance targeted integration significantly (Choulika et al., 1995). Targeted integration (gene knock in) occurs through homology directed repair (HDR)( (Bibikova et al., 2002). In order to enable gene editing by HDR, a DNA ‘;donor’; or repair template containing the desired sequence must be delivered to the cell along with gRNA and Cas9.

Components

Each kit consists of:one vial of gRNA plasmid - KRASone vial of Cas9 GFP plasmidone vial of 120 base donor oligonucleotideone vial of forward PCR primerone vial of reverse PCR primerone vial of genomic DNA control

Features and Benefits

The CRISPR Integration Assay provides a validated CRISPR that cleaves DNA with high efficiency along with a standardized oligo donor designed to survey homology dependent repair frequencies in a wide variety of cell lines CRISPR-aided integration increases the targeted mutagenesis rate without the need for antibiotic selection Integration efficiencies estimated with this kit enable rational design of genome editing experiments in a wide range of cell lines

General description

The CRISPR Integration Kit provides the essential genome editing reagents necessary to integrate a BstNI restriction site to the human KRAS locus. This kit is intended to serve as a tool to evaluate cell lines for ability to integrate new sequences at CRISPR-effected double strand breaks with the aid of short oligonucleotide donor. Integration efficiencies obtained with this kit enable rational design of genome editing experiments in a wide range of cell lines.

Legal Information

CRISPR Use License Agreement