Since its inception in 2012, CRISPR/Cas9 gene editing technology has rapidly become an innovative technology in the research and medical world. This technology is expected to be used to treat genetic diseases such as Cystic Fibrosis, Muscular Dystrophy and Hemophilia. However, how to efficiently deliver the gene-edited Cas9 protein and Guide RNA (sgRNA) into the nucleus remains an important challenge limiting the effectiveness of the CRISPR/Cas9 technology.
A few days ago, researchers at the University of Massachusetts at Amherst developed a new technology that uses nanoparticles to transport Cas9 proteins and sgRNA into cells. The results show that this technology has a 90% success rate of introducing Cas9 protein and sgRNA into cells!
â–² article communication author Professor Vincent Rotello (Source: University of Massachusetts official website)
In a study published in the journal ACS Nano, researchers at the laboratory of the University of Massachusetts Department of Chemistry, Professor Vincent Rotello, modified the Cas9 protein. The Cas9 protein is a highly positively charged protein that does not readily bind autonomously to positively charged pure gold nanoparticles. The researchers added different lengths of glutamate polypeptide chains at the N-terminus of Cas9 protein, which caused local negative domains in Cas9 protein and promoted the binding of Cas9 protein to pure gold nanoparticles. The C-terminus of the newly designed Cas9 protein also incorporates a Nuclear Localization Sequence to help the protein enter the nucleus for gene editing. The researchers called these redesigned Cas9 proteins Cas9En. The complex composed of Cas9En/sgRNA and nanoparticles can fuse with the cell membrane upon contact with the cell membrane and release Cas9En/sgRNA directly into the cytoplasm.
â–²New CRISPR/Cas9En technology (Source: genengnews)
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