A single-guide RNA molecule (sgRNA) is a key component for the CRISPR-Cas9 genome-editng technology, which is one of the most commonly used tools in biological research and has been pursued as a therapeutic tool for treating disease. Previous sgRNA design templates knocked out the target gene with low efficiency, lessening the efficacy of gene knockout therapies.
This new method improves the knockout efficiency of CRISPR-Cas9 technology to such a degree that it is able to efficiently delete entire genes by sgRNA-targeting of two separate loci with high targeting efficiency, thereby eliminating an entire DNA segment from the chromosome. The effectiveness of the technology was demonstrated using a knockout cell line without stable transfection. These data showed that the technology, with slight optimization, was able to achieve more than 95% knockout efficiency in a cell line with transient transfection. This technology is being actively pursued as a therapeutic tool for treating HIV.
- Biological research
- Pharmaceutical research
Features, Benefits & Advantages:
- New sgRNA design template
- Increased knockout efficiency
A Provisional Application Serial 62/216,463 was filed on 09/10/2015.
This technology has been produced and tested.
Haoquan Wu, Biomedical Sciences, Texas Tech University Health Sciences Center El Paso, El Paso, TX.
Keywords: sgRNA, CRISPR, Cas9