dCas9 Protein Allele Sequestration for Improved CRISPR Gene Editing | Rutgers University Innovation Ventures

dCas9 Protein Allele Sequestration for Improved CRISPR Gene Editing

Invention Summary:

CRISPR technology is widely used in research to edit genes in animals and cell lines and is moving towards therapeutic use in the clinic. Inherent to almost all methods employed in gene editing is the lack of control in target gene usage. A method to direct cleavage of only a single target within a cell would be beneficial to research and therapeutic gene editing for the following reasons:

  • 25-30% of gene targets are essential for embryo development, most of which are also necessary for cell growth
  • strong cellular DNA damage response can alter cellular physiology and compromise stem cell integrity

Thus, there is a risk of low success rates for some in vivo CRISPR gene editing projects of both cellular and animal research models and therapeutic editing of stem cells.

Rutgers scientists have developed a novel method, dasCRISPR, that incorporates use of nuclease deficient Cas9 (dCas9), or any similar nuclease deficient Cas-like protein, to allow for modification of a single target allele. This method improves upon and allows for use of CRISPR technology to modify a single allele in cases where modification of both alleles produces deleterious mutations which could adversely affect the survival of the cell/embryo or alter stem cell properties through sustained DNA damage response.


•Allows for editing of genes essential for survival and development in cell and animal models.
•This method is more precise by allowing for protection of one allele of a gene to reduce chance of adverse mutation or downstream effects.
•Allows for greater control over Cas9 gene editing and improves the success rate of CRISPR projects.
•The use of dasCRISPR may also be expanded into non-animal research to generate cell lines.

Market Applications:

•Production of cell lines and animal models when knockout of a single allele is desired, especially when editing of essential genes.
•Controlling therapeutic gene editing by sequestering one allele, thereby moderating the cellular response to unnecessary overediting and/or blocking off-target sites.

Intellectual Property & Development Status: Patent pending. Available for licensing and/or research collaboration. Method fully developed and tested.

Patent Information: