Engineered Guide RNA as a Tool for CRISPR/Cpf1 Gene Editing

This technology aims to optimize the CRISPR-Cpf1 complex by chemically modifying the crRNA nucleotides and engineering crRNA sequences.

The Need

CRISPR technology is a new and exciting tool for editing the genomes of live organisms. With CRISPR, researchers have replaced target genes with chosen alternative genes in species ranging from bacteria to live mice. The CRISPR system uses either the Cas9 or Cpf1 protein to bind a given DNA sequence and cleave it. The CRISPR/Cas9 system is the popular and widely used gold standard, but has some limitations relative to CRISPR/Cpf1. Cas9 requires two RNA molecules to activate DNA cleavage, whereas Cpf1 requires only one guide RNA (crRNA). The Cpf1 crRNA is smaller and less complex than that for Cas9, making some applications easier with Cpf1. Cleavage of DNA with Cas9 results in blunt ends, whereas Cpf1 produces staggered DNA breaks.

The Technology

Researchers in the lab of Dr. Yizhou Dong have invented novel artificial crRNA modifications that increase the efficiency and specificity of naturally-occurring CRISPR/Cpf1. This technology will allow researchers the flexibility of the CRISPR/Cpf1 system while drastically increasing the specificity of cleavage with only a single synthetic RNA.

Commercial Applications

  • Genetics, genomics, and molecular biology reserach
  • Animal disease model generation
  • Cell therapy for diseases such as cystic fibrosis, Duchenne muscular dystrophy, and Fanconi anemia
  • Antimicrobial, antiviral, and biological control (such as mosquito control) applications
  • Genetic modification for agricultural applications
  • Engineering of industrial bacteria, yeast, and fungi for chemical manufacturing

Benefits/ Advantages

  • Requires only one guide RNA
  • Smaller and less complex crRNA
  • More specific targeting
  • More genetic modification possibilities

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