Compositions and methods for purifying polyribonucleotides
AssigneeFlagship Pioneering Innovations VI, LLC
InventorsAlexandra Sophie DE BOER, Nicholas McCartney PLUGIS, Catherine CIFUENTES-ROJAS, Ki Young PAEK, Elissa Magdalene HOBERT, Joshua Nathan FARB, Vadim DUDKIN
Life of this patent
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Abstract
The present disclosure relates to compositions and methods for separating and/or purifying polyribonucleotides. The polyribonucleotide may be separated from a mixture of polyribonucleotides with an oligonucleotide that hybridizes to a target region of the polyribonucleotide.
Claims60
A method of separating a linear polyribonucleotide from a plurality of polyribonucleotides comprising a mixture of linear polyribonucleotides and circular polyribonucleotides comprising an open reading frame (ORF) encoding a polypeptide, the method comprising:
- (a) providing a sample comprising the plurality of polyribonucleotides, wherein a subset of the plurality of polyribonucleotides comprise the linear polyribonucleotide;
- (b) attaching a target region to the linear polyribonucleotide;
- (c) contacting the sample with an oligonucleotide that hybridizes to the target region;
- and
- (d) separating the linear polyribonucleotide comprising the target region that is hybridized to the oligonucleotide from the plurality of polyribonucleotides in the sample.
A method of separating a linear polyribonucleotide from a plurality of polyribonucleotides comprising a mixture of linear polyribonucleotides and circular polyribonucleotides, the method comprising:
- (a) providing a sample comprising the plurality of polyribonucleotides, wherein a subset of the plurality of polyribonucleotides comprise the linear polyribonucleotide;
- (b) attaching a target region to the linear polyribonucleotide;
- (c) contacting the sample with a column comprising a resin comprising a plurality of particles conjugated to an oligonucleotide that hybridizes to the target region;
- and
- (d) collecting an eluate comprising a portion of the sample that is not hybridized to the oligonucleotide from the plurality of polyribonucleotides in the sample.
A method of separating a linear polyribonucleotide from a plurality of polyribonucleotides comprising a mixture of linear polyribonucleotides and circular polyribonucleotides, the method comprising:
- (a) circularizing a linear precursor to form the circular polyribonucleotide;
- (b) providing a sample comprising the plurality of polyribonucleotides, wherein a subset of the plurality of polyribonucleotides comprise the linear polyribonucleotide;
- (c) attaching a target region to the linear polyribonucleotide;
- (d) contacting the sample with an oligonucleotide that hybridizes to the target region;
- and
- (e) separating the linear polyribonucleotide comprising the target region that is hybridized to the oligonucleotide from the plurality of polyribonucleotides in the sample.
Dependencies are inferred from the claim text.