Rna affinity purification
AssigneeModernaTX, Inc.
InventorEdward J. Miracco
Life of this patent
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Abstract
Provided herein, in some embodiments, are methods of purifying a nucleic acid preparation. The methods may comprise contacting a nucleic acid preparation comprising messenger ribonucleic acid with an RNase III enzyme that is immobilized on a solid support and binds to double-stranded RNA contaminants.
Claims35
A method of purifying a nucleic acid preparation, comprising
- contacting a nucleic acid preparation comprising messenger ribonucleic acid (mRNA) with an RNase III enzyme that is immobilized on a solid support, under conditions that result in binding of the RNase III enzyme to double-stranded RNA.
The method of claim 14, wherein the 5′ terminal cap is 7mG(5′)ppp(5′)NlmpNp.
A method, comprising:
- performing an in vitro transcription reaction in the presence of a template nucleic acid to produce an in vitro transcription product;
- and
- contacting the in vitro transcription product with an RNase III enzyme that is immobilized on a solid support.
A composition comprising RNA purified according to a method that comprises contacting a preparation comprising messenger ribonucleic acid (mRNA) with an RNase III enzyme that is immobilized on a solid support.
A composition comprising RNA prepared according to a method that comprises
- performing an in vitro transcription reaction in the presence of a template nucleic acid to produce an in vitro transcription product;
- and
- contacting the in vitro transcription product with an RNase III enzyme that is immobilized on a solid support.
A method of purifying a product of an in vitro transcription reaction, comprising
- contacting a product of an in vitro transcription reaction comprising messenger ribonucleic acid (mRNA) with a catalytically inactive thermostable RNase III enzyme that is immobilized on a solid support, under conditions that result in binding of the RNase III enzyme to double-stranded RNA.
A catalytically inactive thermostable RNase III enzyme having an amino acid sequence identified by SEQ ID NO:
- 4.
Dependencies are inferred from the claim text.