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Research Update

Cell Specific Snipping with miRNA-Cas9 Switches

Recently, a group of Japanese researchers published work that incorporated TriLink’s modified NTPs into custom Cas9 mRNA to optimize cell specific applications for CRISPR Cas9 gene editing.

Although DNA based gene editing has evolved rapidly, particularly for CAR-T therapeutics (Novartis’ Kymriah and Gilead’s Yescarta), one issue limiting its success in some applications is the ability to function in a cell-type specific manner. The Research Team opted for an RNA-based delivery system, which circumvents several of the safety issues associated with DNA-based systems, including genome integration.

"In order to regulate the endonuclease activity of Cas9, several methods have been reported, including optogenetic or small molecule/protein-inducible systems, a tissue-specific promoter system, and transcriptional regulatory circuits using split-Cas9. However, these reports have predominantly used DNA delivery approaches," the researchers reported.

The Team engineered an mRNA-based Cas9 switch that is modulated by cell-type specific microRNA (miRNA). Specifically, they introduced miRNA-complementary sequences into the 5’UTR of Cas9 mRNA, hypothesizing that this could selectively control genome editing by sensing endogenous miRNA levels within a heterogeneous cell population...

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    What base modifications can be incorporated into my custom mRNA?

    TriLink offers a variety of modified rNTPs suitable for in vitro transcription. We have assessed transcription efficiency based on final product formation of a 1.9 kb transcript with the following triphosphates at 100% substitution using...

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