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US 12,612,658 B2Granted
RNA replication using transcription polymerases
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsNimit Jain, Andrew Z. Fire, Lucas R. Blauch, Sindy K. Y. Tang, Karen L Artiles, Julia T. Garcia-Daou, Y. Whitney Yin, Michal R. Szymanski
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Abstract
Compositions and methods for amplifying RNA by replication using transcription polymerases are disclosed. Such replicated RNAs can be used in various applications such as RNAi therapeutics, diagnostic probes, RNA sequencing, directed evolution of RNA aptamers without intermediate conversion to DNA, and RNA vaccines. The transcription polymerases comprise T7 bacteriophage RNA polymerase.
Claims18
A method of amplifying RNA comprising:
- constructing an RNA template for said amplifying the RNA, the RNA template comprising (i) a 2-way repeat configuration comprising a first inverted repeat, (ii) a 4-way repeat configuration comprising a second inverted repeat that is shorter than the first inverted repeat, wherein each arm of the 2-way repeat comprises the second inverted repeat, (iii) a 3′ end sequence comprising GG or CC and one or more additional nucleotides following the GG or the CC, and (iv) a nucleotide sequence of interest;
- and
- replicating the RNA in a reaction mixture comprising:
- an RNA polymerase;
- a set of ribonucleoside triphosphates comprising ATP, CTP, GTP, and UTP, or analogues or derivatives thereof;
- and
- the RNA template.
4 more elements
Dependencies are inferred from the claim text.