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The first in-house CRISPR application to design both gRNA and donor template sequences required for genome editing. is a complete and fully customizable tool to facilitate and optimize your CRISPR experiments. © The introduction of targeted genomic sequences changes by CRISPR technology into living cells is becoming a powerful tool for gene therapy or disease modelling. CRISPR only requires a nuclease and customized nucleic sequences. Preliminary bioinformatics analysis for both gRNA design and donor template can improve the success of the experiment. This is where the CRISPR LifePipe will make genome editing as simple as using a text editor. DESIGN OF RNA GUIDE (OR gRNA).. The RNA guide, or gRNA, is a short RNA sequence which guides the Cas9 endonuclease to the targeted region to cut on the genome. gRNA is crucial for CRISPR gene editing because it provides targeting efficiency on the region of interest on the genome while limiting the non-specific off-targets. Our gRNA design tool is built to allow you to target different genomic regions: • Targeting a region present on most of the transcripts of a gene • Targeting an exon or an intron of a transcript • Targeting UTR region of a gene • Targeting a particular amino acid CRISPR technology is a genome engineering technology which allows accurate genome modification of a cell. It is based on the immune system CRISPR/Cas9 discovered in bacteria. Cho et al. Nat Biotech 2013 Our tool selects gRNA sequences with the lowest number of off-targets to avoid side effects within the cell. It also designs efficient gRNA estimating the gRNA secondary structures and calculating the number of SNPs on its sequence. Advanced options provide a precise configuration to optimize the design of gRNA. Indeed the tool offers different specific designs parameters like the GC percent of the sequence, the presence of TLR motifs or the presence of nucleodide stretches on the RNA sequence. Cong et al. Science 2013 DESIGN OF DONOR TEMPLATE.. CRISPR technology is able to modify specific DNA sequence using a RNA guide (gRNA). This short RNA sequence guides the Cas9 endonuclease to the genomic region to target and cut. A donor template sequence uses DNA repair mecanism of the cell to allow modifications of DNA sequence. The donor template is a DNA sequence inserted into the cell along with the gRNA and the Cas9 endonuclease. It is acting by homologous recombination during DNA repair mecanism to replace DNA sequence of the cell by the modified DNA sequence of the donor template. • Insertion of a mutation into a gene • Gene tagging in 5’ or 3’ “ CRISPR: GENE EDITING IS JUST THE BEGINNING The real power of the biological tool lies in exploring how genomes work” Nature, 2016 • Gene Knock-Out Our tool can design plasmid donor sequences with variable lengths of the two homology arms and different mutations or tag to insert into the cell, or ssDNA oligo (ssODN). An option allows the insertion of a particular DNA fragment between the two homology arms, like a selection cassette. The gRNA is always inactivated by our tool to prevent the Cas9 to cut the donor sequence. A quality control of the donor template is also systematically performed. The application returns a graphical output to visualize the full donor template sequence, the position of the gRNA sequence and the inserted variations on the donor. WHY CHOOSE CRISPR LIFEPIPE?.. The CRISPR tools have been developped by Life&soft team to meet all the needs of the CRISPR users. These tools will facilitate and improve all steps required for a high quality and successful CRISPR experiment preparation. Convenient Our application gather two different tools together required for CRISPR genome editing, from gRNA design to the choice of donor template sequence Complete Our tool has been developped considering the new findings and hints published into scientific literature. Supported Both tools are updated and supported by our team regularly. Flexible Easy to use All our CRISPR tools are entirely customizable regarding your specific and evolving needs : species to study, PAM and Cas9 used… Intuitive web interface fully customizable by user regarding his specifc needs. Currently available on BIOVIA Pipeline Pilot®. The portability of the Easy to deploy application out-of-the-box on linux environment is being developped. CONTACT US.. Les Colocs (Parc Noveos) 8 B avenue Descartes - 92350 Le Plessis-Robinson - France [email protected] +33 (0) 6 19 53 89 27 www.lifeandsoft.com www.chrysalis-lamarque.fr - Photos & graphiques : Shutterstock.com - Fotolia.com - Life&Soft - Servier Medical Art de Servier est mis à disposition selon les termes de la licence Creative Commons Attribution 3.0 France Different donor sequences can be designed with our tool regarding the modification that is expected in the cell: