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US 2024/0240218 A1Published application
Methods of enriching for circular polyribonucleotides
AssigneeFlagship Pioneering Innovations VI, LLC
InventorsAlexandra Sophie De Boer, Elissa Magdalene HOBERT, Joseph Arthur DEPETER
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Abstract
The methods of the disclosure can be used to enrich a population of circular polyribonucleotides in a mixture of linear polyribonucleotides and circular polyribonucleotides.
Claims48
A method of producing an enriched population of circular polyribonucleotides, the method comprising:
- (a) providing a population of polyribonucleotides comprising circular polyribonucleotides and linear polyribonucleotides;
- (b) digesting at least a portion of the linear polyribonucleotide with a 5′ exonuclease in an amount between 0.01 U and 0.2 U per 1 μg of polyribonucleotides;
- and
- (c) digesting at least a portion of the linear polyribonucleotide with a 3′ exonuclease in an amount between 0.4 U and 4 U per 1 μg of polyribonucleotides;
- thereby producing an enriched population of circular polyribonucleotides.
2 more elements
A method of producing an enriched population of circular polyribonucleotides, the method comprising:
- (a) providing a population of polyribonucleotides comprising circular polyribonucleotides and linear polyribonucleotides;
- (b) digesting at least a portion of the linear polyribonucleotide with a 5′ exonuclease, and
- (c) digesting at least a portion of the linear polyribonucleotide with a 3′ exonuclease,
- wherein digesting steps (b) and (c) are performed in a digesting buffer comprising between 0.05 mM and 1 mM Mg2+ and/or between 0.1 mM and 5 mM dithiothreitol;
- thereby producing an enriched population of circular polyribonucleotides.
2 more elements
Dependencies are inferred from the claim text.