Complete Cpf1-based CRISPR genome editing system

Integrated DNA Technologies (IDT) is the first genomics company to develop and bring to market a complete ribonucleoprotein-based Cpf1 CRISPR system.

The Alt-R A.s. Cpf1 CRISPR System inherits the traits of IDT’s Cas9-based system while taking advantage of Cpf1’s natural AT-rich target sequence preference and ability to make staggered cuts.

In addition, IDT is launching an associated range of CRISPR support tools to expand experimental options and capabilities for molecular biology researchers.

The new tools extend the ease-of-use and performance of IDT’s Alt-R system through options for fluorescent visualisation, enhanced nuclease transfection, and genome editing detection.

Together, the new expanded Alt-R range breaks barriers to wider target spaces not addressable by Cas9 systems alone, and provides a level of flexibility in experimental design not previously possible.

CRISPR is now one of the most widely used tools for genome modification. IDT’s Alt-R System already overcomes the limitations of using sgRNAs in the ribonucleoprotein (RNP) complex by enhancing editing efficiency and lowering toxicity.

Now, in developing a complementary Cpf1-based system, IDT has opened up options for targeting AT-rich sequences. The new system includes the Alt-R A.s. Cpf1 nuclease, containing two integrated nuclear localisation sites, which complexes as an RNP with a minimal 41-44 nucleotide Alt-R CRISPR-Cpf1 crRNA. The system requires no tracrRNA, reducing potential reagent costs and experimental complexity.

The expanded range of Alt-R support tools now includes the Alt-R CRISPR-Cas9 tracrRNA—5’ ATTO 550, and Alt-R Electroporation Enhancers for Cas9 and Cpf1.

The former allows in vivo fluorescent visualisation of the RNP complex and FACS enrichment of transfected cells without affecting RNP functionality, while the enhancers improve the efficiency of transfection using the Nucleofector (Lonza) and Neon (Thermo) electroporation systems. These new tools enable researchers to improve the overall efficiency of their genome editing over traditional methods.

In addition, the new Alt-R Genome Editing Detection Kit provides a fast, easy, and low-cost method for confirming editing events with a T7 Endonuclease I (T7EI)-based mismatch detection system. These additions to the Alt-R range, combined with the new Cpf1 system, provide researchers with cost-effective and reliable solutions for performing and evaluating CRISPR genome editing experiments.

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