integrations, User guides & BV3L6 Cas12a (Cpf1), called Cas12a Ultra. The cleavage site is specified by the spacer element of the crRNA (blue bar). The new mutant enzyme has enhanced editing activity, reaching or exceeding the performance of Cas9. Solutions for your research needs. The new mutant enzyme has enhanced editing activity, reaching or exceeding the performance of Cas9. Cas9 and Cas12a reagents are available from IDT. New Alt-R A.s. Cas12a (Cpf1) Ultra exhibits increased genomic editing efficiency in Jurkat and HEK-293 cells. : Our scientists have computationally designed and tested negative control crRNAs to be non-targeting in human, mouse, and identify locations in your target region with the protospacer adjacent motif (PAM) sequence, TTTV, where V is A, C, or genomics, GMP, OEM & integrations, User guides & other Cas enzymes, RNP complexes (4 μM) were delivered into Jurkat and HEK-293 cells via a Nucleofector™ system (Lonza) Its activity is independent of the enzyme employed; IDT’s new CRISPR Cas12a (Cpf1) Ultra enzyme can target new sites within the genome and with greater efficiency. Alt-R Predesigned Cas9 crRNA Selection Tool, Order status (Genes & gene fragments only), Target Capture Probe Design & Ordering Tool, single guide RNA (sgRNA) or a 2-part guide RNA, complexing the enzyme and gRNA to form a ribonucleoprotein (RNP), CRISPR genome in addition to TTTV motifs, increasing target range for genome editing studies. CRISPR-Cas12a genome editing method uses the Cas12a endonuclease to generate double-stranded breaks that contain a staggered 5′ overhang. This versatile reagent is also compatible with electroporation and lipofection methods. protocols, Safety data The new tool will find equal utility for both knock-out as well as knock-in applications, in fields ranging from basic research cell engineering to medical gene therapy to agricultural biotechnology. The 2 main Cas enzymes used in genome editing are Cas9 and Cas12a, Cas9 was the first Cas enzyme used in the earliest CRISPR genome editing projects, and it remains the most popular Cas enzyme. sites. generation sequencing, Genes & Alt-R Predesigned Cas9 crRNA Selection Tool, Order status (Genes & gene fragments only), Target Capture Probe Design & Ordering Tool, Alt-R CRISPR-Cas12a (Cpf1) genome editing, CRISPR-Cas12a (Cpf1) crRNA Oligo Entry tool, Alt-R CRISPR-Cpf1—RNP electroporation, Amaxa Nucleofector system, Alt-R CRISPR-Cpf1—RNP electroporation, Neon Transfection system, Reference protocol: Perform gene knockout with the Alt-R CRISPR-Cas System. Improved enzymes:  The Alt-R Cas12a (Cpf1) enzyme has recently been further optimized to deliver even higher performance. IDT developed a novel mutant of the Acidaminococcus sp. Thomasnet Is A Registered Trademark Of Thomas Publishing G. Your Alt-R CRISPR-Cas12a (Cpf1) crRNA will bind to the DNA strand opposite to the PAM sequence (Figure 2). Common incorrect design examples are shown as crossed-out gray text. design checker tool to assess targeting potential before ordering Cas9 gRNAs that are synthesized using Alt-R gRNA modifications. Sample data from T7 endonuclease I (T7EI) assays of samples transfected with Cas12a HPRT Positive Controls. The Alt-R CRISPR-Cas9 System is an optimized genome editing solution that outperforms other CRISPR approaches for producing on-target, double-stranded DNA breaks. sequence in the Cpf1 crRNA ordering page: Rat Hprt1, Cas12a Positive Control crRNA: ACCGCCCCCCCCATACCCCAA. Negative control crRNAs are important for showing that transfection of the RNP complex is not responsible for observed phenotypes. Next generation sequencing method for sequencing on-target and known off-target CRISPR editing sites. Tool, gBlocks Gene IDT’s Alt-R Cas12a (Cpf1) Ultra is a first addition to the genome editing toolbox that offers the same efficiency and versatility as Cas9. Cell lines used were HEK-293 (human), Hepa1-6 (mouse), and RG2 (rat). result does not rule out off-target effects of your experimental crRNA. editing, Next For designs of your own or from publications, use our The new mutant enzyme has enhanced editing activity, reaching or exceeding the performance of Cas9. Publication of the Cas12a system in 2015 generated considerable interest among researchers seeking new enzymes for genome editing experiments, particularly due to its unique PAM site (“TTTV”) that greatly expanded the number of potential editing sites in the genome beyond that of Cas9 (“NGG”). Plates require a minimum order of 24 crRNAs. Integrated DNA Technologies, Inc. (IDT) develops, manufactures, and markets nucleic acid products for the life sciences industry in the areas of academic and commercial research, agriculture, medical diagnostics, and pharmaceutical development. Enlist Your Company ico-arrow-default-right. 5 µM of Alt-R Cas12a RNP, together with 3 µM of Alt-R Cas12a Electroporation Enhancer and 3 µM of single-stranded DNA template (Ultramer oligos, If your company can help provide supplies, capabilities, or materials for products such as N-95 Masks and Tyvek Suits — Please let us know. The latest version (Ultra) can be directly substituted into this protocol in place of the prior Alt-R Cas12a V3 enzymes. HDR required RNP and templates (i.e., donor oligos containing the insertion sequence) and can be used to knockin specific mutations (e.g., insertions, deletions, SNP mutations). Ribonucleoprotein (RNP) complexes were formed with wild type (WT) or Alt-R A.s. Cas12a (CPF1) Ultra (Ultra), A “no treatment” control group that received media alone was also included in the experiment (dark blue bar). To deliver Cas enzyme and guide RNA to cells, we recommend complexing the enzyme and gRNA to form a ribonucleoprotein (RNP). We make them better, and deliver them faster. Editas Medicine, a leading company developing genome editing medicines, recognized the potential advantages of the newly discovered Cas12a (also known as Cpf1) system and obtained an exclusive license from the Broad Institute, Inc., acting on behalf of itself and the President and Fellows of Harvard College, the Massachusetts Institute of Technology, Wageningen University, and the University of Tokyo in 2016 for human therapeutics.

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