Artificial nucleic acid molecules for improved protein expression
AssigneeCureVac AG
InventorsAndreas Thess, Thomas Schlake, Stefanie Grund
Life of this patent
Estimating the term…
Abstract
The invention relates to an artificial nucleic acid molecule comprising an open reading frame and a 3′-UTR comprising at least one poly(A) sequence or a polyadenylation signal. The invention also relates to a method for increasing protein production from an artificial nucleic acid molecule and to the use of a 3′-UTR for a method for increasing protein production from an artificial nucleic acid molecule. Moreover, the invention concerns the use of the artificial nucleic acid molecule as a medicament, as a vaccine or in gene therapy.
Claims28
A method for stimulating an immune response in an organism, the method comprising administering to the organism a RNA molecule comprising:
- a) a 5′-cap structure;
- b) an open reading frame encoding an antigen;
- and
- c) a heterologous 3′-untranslated region comprising a first and a second poly(A) sequence, wherein:
- (i) the first poly(A) sequence comprises at least 20 adenine nucleotides;
- and
- (ii) the second poly(A) sequence comprises at least 70 adenine nucleotides,
- wherein the first and the second poly(A) sequences are separated by a nucleic acid sequence comprising from 10 to 90 nucleotides and having no more than 2 consecutive adenine nucleotides,
- wherein when the RNA molecule is expressed in the organism, the RNA molecule yields increased expression of the antigen encoded by the open reading frame in comparison to a reference nucleic acid molecule comprising an identical nucleic acid sequence as the RNA molecule but lacking the second poly(A) sequence;
- and
- wherein the RNA molecule comprises at least one nucleotide analog comprising a modified form of uridine chemically altered by methylation.
A method for stimulating an immune response in an organism, the method comprising administering to the organism a RNA molecule comprising:
- a) a 5′-cap structure;
- b) an open reading frame encoding an antigen;
- and
- c) a heterologous 3′-untranslated region comprising a first and a second poly(A) sequence, wherein:
- (i) the first poly(A) sequence comprises at least 30 adenine nucleotides;
- and
- (ii) the second poly(A) sequence comprises at least 70 adenine nucleotides,
- wherein the first and the second poly(A) sequences are separated by a nucleic acid sequence comprising 10 nucleotides and having no more than 2 consecutive adenine nucleotides,
- wherein when the RNA molecule is expressed in the organism, the RNA molecule yields increased expression of the antigen encoded by the open reading frame in comparison to a reference nucleic acid molecule comprising an identical nucleic acid sequence as the RNA molecule but lacking the second poly(A) sequence;
- wherein the RNA molecule is formulated with a cationic lipid;
- and
- wherein the RNA molecule is administered intramuscularly.
A method for stimulating an immune response in an organism, the method comprising administering to the organism a RNA molecule comprising:
- a) a 5′-cap structure;
- b) an open reading frame encoding a coronavirus antigen;
- and
- c) a heterologous 3′-untranslated region comprising a first and a second poly(A) sequence, wherein:
- (i) the first poly(A) sequence comprises at least 20 adenine nucleotides;
- and
- (ii) the second poly(A) sequence comprises at least 70 adenine nucleotides,
- wherein the first and the second poly(A) sequences are separated by a nucleic acid sequence comprising 10 nucleotides and having no more than 2 consecutive adenine nucleotides,
- wherein the RNA molecule is administered intramuscularly;
- wherein when the RNA molecule is expressed in the organism, the RNA molecule yields increased expression of the antigen encoded by the open reading frame in comparison to a reference nucleic acid molecule comprising an identical nucleic acid sequence as the RNA molecule but lacking the second poly(A) sequence;
- wherein the RNA molecule comprises at least one nucleotide analog comprising a modified form of uridine chemically altered by methylation;
- and
- wherein the RNA molecule is transfected into cells of the organism in a nanoparticle.
A method for stimulating an immune response in an organism, the method comprising administering to the organism a RNA molecule comprising:
- a) a 5′-cap structure;
- b) an open reading frame encoding a coronavirus antigen;
- and
- c) a heterologous 3′-untranslated region comprising a first and a second poly(A) sequence, wherein:
- (i) the first poly(A) sequence comprises at least 30 adenine nucleotides;
- and
- (ii) the second poly(A) sequence comprises at least 70 adenine nucleotides,
- wherein the first and the second poly(A) sequences are separated by a nucleic acid sequence comprising 10 nucleotides and having no more than 2 consecutive adenine nucleotides,
- wherein the RNA molecule is administered intramuscularly;
- wherein when administered intramuscularly, the RNA molecule yields an increased neutralizing antibody response in comparison to an intramuscular injection of a reference nucleic acid molecule comprising an identical nucleic acid sequence as the RNA molecule but lacking the second poly(A) sequence;
- wherein the RNA molecule comprises at least one nucleotide analog comprising a modified form of uridine chemically altered by methylation;
- and
- wherein the RNA molecule is transfected into cells of the organism in a nanoparticle.
Dependencies are inferred from the claim text.