CRISPR/Cas9 is a simple and efficient genome editing tool. Although gene knockout cell lines can be generated by gRNAs without donor vector, the screening process can be very tedious. OriGene offers genome-wide CRISPR gene knockout / knockin kits containing 2 gRNA vectors and donor DNA. The selection marker in the donor cassette greatly facilitates the screening process.

Two types of CRISPR knockout kits are offered, differing in repair mechanism that is used to achieve gene knockout, homology based or non-homology based. HDR-mediated knockout kit is our first generation gene knockout kits, gene knockout is achieved via homology directed repair (HDR). KN2.0 gene knockout kit is an improved version, gene knockout is achieved via non-homology based repair. KN2.0 CRISPR knockout kit is more efficient and works in both dividing and non-dividing cells.

  1. Non-homology mediated CRISPR gene knockout kits (KN2.0) (Validation Data)
  2. HDR mediated CRISPR gene knockout kits (Validation Data)

Comparison of CRISPR gene knockout kits

 

HDR mediated KN2.0 non-homology mediated
gRNA vectors pCas-Guide pCas-Guide
Donor Donor cassette flanking with homologous arms Donor cassette without homologous arms
Cell spectrum Dividing cells Dividing and non-dividing cells
Knockout Efficiency Medium High

 

KN2.0 non-homology mediated CRISPR gene knockout kits

Gene knockout is based on non-homology-mediated repair mechanism. After gRNA targeted double stranded DNA cleavage, the linear donor DNA containing a selection cassette will be integrated at the gRNA cutting site at forward or reverse direction. Our testing data using KN2.0 kits showed that the vast majoiry gene knockout is bioallelic, one allele contains donor integration, the other allele has indels (insertion and deletion), which might change protein coding or cause premature stop.

 

Features

  • Ready-to-use gene knockout kit
  • 2 guide RNA vectors in pCas-Guide
  • Donor cassette, EF1a-GFP-P2A-Puro
  • Non-homology mediated gene knockout
  • Highly efficient gene knockout kit

Scheme of gene knockout of KN2.0