Editas Medicine. has filed a patent for compositions, systems, and methods for analyzing the activity of nucleases. The patent claims a method involving emulsifying a library of nucleic acid templates, expressing RNA-guided nuclease variants, subjecting the droplets to conditions for nuclease cleavage, ligating the cleaved templates with capture probes, and identifying the RNA-guided nuclease variants. GlobalData’s report on Editas Medicine gives a 360-degree view of the company including its patenting strategy. Buy the report here.
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According to GlobalData’s company profile on Editas Medicine, CRISPR genome editing was a key innovation area identified from patents. Editas Medicine's grant share as of September 2023 was 14%. Grant share is based on the ratio of number of grants to total number of patents.
Method for analyzing activity of nucleases using droplet emulsification
A recently filed patent (Publication Number: US20230313174A1) describes a method for identifying RNA-guided nuclease variants using emulsification and ligation techniques. The method involves emulsifying a library of nucleic acid templates, each containing a first nucleotide sequence encoding a variant of an RNA-guided nuclease and a second nucleotide sequence with a target site for the nuclease. This emulsification process forms droplets, with each droplet containing a unique nucleic acid template. The RNA-guided nuclease variants are then expressed within the droplets. The droplets are subjected to conditions that promote nuclease cleavage of the target site, resulting in a population of cleaved nucleic acid templates. These cleaved templates are then ligated with oligonucleotide capture probes specific for the cleaved ends, producing a variety of ligation products. The method further includes the identification of at least one first nucleotide sequence encoding an RNA-guided nuclease variant within the ligation products.
In addition to the above steps, the patent also mentions the disruption of the droplets before the ligation step to obtain a mixture of cleaved nucleic acid templates. The method may also involve the detection, amplification, or sequencing of the ligation products. Each nucleic acid template in the library may contain a third nucleotide sequence encoding a guide RNA, which can be linked to the first promoter or a second promoter. Furthermore, the nucleic acid templates may contain a fourth nucleotide sequence adjacent to the target site, comprising a protospacer adjacent motif (PAM). The predetermined cleaved end of the cleaved nucleic acid templates can be a 5' phosphate group, a blunt end, or a cohesive end with a 3' overhang or a 5' overhang with a specific number of nucleotides. The ligating step can involve the use of T4 ligase or E. coli ligase. Multiple oligonucleotide capture probes can be used for ligating the cleaved nucleic acid templates, with each probe specific for a different predetermined cleaved end. These probes may also contain unique detectable labels such as barcode sequences or fluorescent markers.
Overall, this patent describes a method for efficiently identifying RNA-guided nuclease variants using emulsification and ligation techniques. The method allows for the generation of a population of cleaved nucleic acid templates and the subsequent identification of specific RNA-guided nuclease variants within these templates. This technique has potential applications in various fields, including genetic engineering and gene editing.
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GlobalData Patent Analytics tracks bibliographic data, legal events data, point in time patent ownerships, and backward and forward citations from global patenting offices. Textual analysis and official patent classifications are used to group patents into key thematic areas and link them to specific companies
