Colorado Patent of the Month – November 2021

The scientific community has long been working towards precise and targeted changes to the genome of living cells. These efforts have identified certain nucleases that allow manipulation of gene sequences and therefore function. The nucleases work well as they include nucleic acid-guided nucleases which allow for these permanent edits to be made in the cell. Inscripta, Inc. is working with these nucleases in an effort to make digital genome engineering a real possibility.

Recently, they have developed a system to edit nucleic acids in live yeast cells. Yeast use gRNA expression systems with RNA polymerase III (pol III) or II (pol II) to drive transcription. Pol III have lower expression levels than pol II, which are more tolerant to sequence variations and allow for increased sequence flexibility. So while pol II sounds like the way to go, they have their own limitations – gRNA’s expressed from the pol II promoter must remain in the nucleus. A self-cleaving ribozyme is used to help cleave off the export tag, allowing the product to meain in the nucleus. 

As such, their system uses a ribozyme-containing editing cassette for nucleic acid-guided nuclease editing. The yeast genome can be edited using this editing cassette and a single vector backbone which ultimately allows for multiple edits per round. 

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