Expanding upon the CRISPR-Cas9 System

news October 29 2015

The CRISPR-Cas9 system has proven to be an indispensable tool for genome editing, improving targeting efficiency and ease of use with the ability to generate single point mutations and even change protein domains.

The CRISPR-Cas9 system has proven to be an indispensable tool for genome editing, improving targeting efficiency and ease of use with the ability to generate single point mutations and even change protein domains.

Recently, researchers have discovered a new CRISPR endonuclease which may be more powerful: Cpf1 derived from bacteria. They found that Cpf1 allows for more flexibility, as it only needs a single RNA, lacks tracrRNA required in Cas9, and targets distinct protospacer-adjacent motif (PAM) sequences farther from the recognition site. Most importantly, using Cpf1 results in DNA that is cleaved with staggered short overhang ends, as opposed to the blunt, mutation-prone ends achieved with Cas9. This improvement allows for the further fine-tuning of CRISPR applications, with Cpf1 capable of editing genomic loci in human cells.

Study authors used the Direct-zol™ RNA MiniPrep from Zymo Research for heterologous E. coli expression of the FnCpf1 locus by means of RNA-seq.

 

7 minutes from TRIzol to purified RNA – Watch the video to learn more

Check out the protocol to learn more! Click here


Zetsche, B., et al. (2015) Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell. 2015.

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