CRISPR Tools

CRISPR Tools: Products and Services for Genome Engineering

Discovery of CRISPR-Cas9 based genome engineering provides a powerful new tool in biology. The prokaryote-derived CRISPR-Cas system allows highly specific genomic modification of your genes of interest, by using a combination of sgRNA (single guide RNA, or gRNA) along with Cas9 nuclease. CRISPR-technology enables knock out, knock in, inhibition or activation of any part of the genome with unprecedented specificity. The ease of use and robustness of CRISPR-Cas9 have revolutionized genome editing for research ranging from fundamental science to translational medicine.

CRISPR Tools: sgRNA Scaffold and Target Sequence Design

Our partner Cellecta have developed an improved sgRNA scaffold showing higher Cas9 binding, called Hybridization Extended A-T Inversion (HEAT) scaffold which improves screening results:

For robust results sgRNA target sequences need to be highly efficient and specific.

Cellecta´s sgRNA efficiencies are determined by using the criteria for effective guides described by Doench, et al. 2016 and Azimuth activity scores are used to rank the guides for efficacy. The designed guides are cross-checked against the UCSC CRISPR targets track to ensure they are unique in the genome. The UCSC specificity quartile is used to score for off-target potential.

Our partner transOMIC are using the CROATAN sgRNA algorithm (Erard et al., 2017, Molecular Cell) which combines a machine-learning approach with other strategies to optimize knockout efficiency with the CRISPR/Cas9 system. In addition a multiplexed sgRNA expression strategy was developed that promotes the functional ablation of single genes and allows for combinatorial targeting. These strategies have been combined to design and construct a genome-wide, sequence-verified, arrayed CRISPR library. This resource allows single-target or combinatorial genetic screens to be carried out at scale in a multiplexed or arrayed format.