CRISPR Integration Kit
SIGMA/HSOLIGOINT
Product Type: Chemical
CEL-I and BstNI cleavage at Kras target site in K562 cell line using CRISPR Integration Assay.
| application(s) | CRISPR genome editing |
| packaging | pkg of (6 vials per kit) |
| Quality Level | 200  |
| recombinant | expressed in E. coli |
| selection | kanamycin |
| shipped in | dry ice |
| storage temp. | −20°C |
| 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. |
| 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 |
| Other Notes: | Each kit consists of: one vial of gRNA plasmid - KRAS one vial of Cas9 GFP plasmid one vial of 120 base donor oligonucleotide one vial of forward PCR primer one vial of reverse PCR primer one vial of genomic DNA control |
| Storage Temp. | −20°C |
| UNSPSC | 12352200 |