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US 11,230,720 B2Granted
Nucleic acid molecules containing spacers and methods of use thereof
AssigneeAudentes Therapeutics, Inc.
InventorJohn T. Gray
Life of this patent
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Abstract
The present invention relates nucleic acid molecules and concatemers containing spacer sequences useful for the efficient packaging of viral particles so as to minimize the incorporation of contaminant nucleic acids into these vectors, as well as methods of producing such viral particles.
Claims56
A nucleic acid molecule comprising:
- (i) a first spacer (SS1);
- (ii) a first inverted terminal repeat (ITR1);
- (iii) a cloning site (CS);
- (iv) a second inverted terminal repeat (ITR2);
- and
- (v) a second spacer (SS2);
- operably linked to each other in a 5′-to-3′ direction as:
- SS1-ITR1-CS-ITR2-SS2, wherein the SS1 and the SS2 each, independently, comprise a portion having a nucleic acid sequence that is at least 85% identical to any one of the nucleic acid sequences of SEQ ID NOS:
- 3-5 and 9-11, and wherein neither the SS1 nor the SS2 comprises an open reading frame that encodes more than 100 amino acids.
6 more elements
A nucleic acid molecule comprising:
- (i) a SS1;
- (ii) an ITR1;
- (iii) a heterologous polynucleotide molecule (HPM);
- (iv) an ITR2;
- and
- (v) a SS2;
- operably linked to each other in a 5′-to-3′ direction as:
- SS1-ITR1-HPM-ITR2-SS2, wherein the SS1 and the SS2 each, independently, comprise a portion having a nucleic acid sequence that is at least 85% identical to any one of the nucleic acid sequences of SEQ ID NOS:
- 3-5 and 9-11, and wherein neither the SS1 nor the SS2 comprises an open reading frame that encodes more than 100 amino acids.
6 more elements
A nucleic acid molecule comprising:
- (i) a SS1;
- (ii) an ITR1;
- (iii) a first homology arm (HR1);
- (iv) a CS;
- (v) a second homology arm (HR2);
- (vi) an ITR2;
- and
- (vii) a SS2;
- operably linked to each other in a 5′-to-3′ direction as:
- SS1-ITR1-HR1-CS-HR2-ITR2-SS2, wherein the SS1 and the SS2 each, independently, comprise a portion having at least 85% sequence identity to a nucleic acid sequence selected from the group consisting of the nucleic acid sequences of SEQ ID NOS:
- 3-5 and 9-11, and wherein neither the SS1 nor the SS2 comprises an open reading frame that encodes more than 100 amino acids.
8 more elements
A nucleic acid molecule comprising:
- (i) a SS1;
- (ii) an ITR1;
- (iii) a HR1;
- (iv) a HPM;
- (v) a HR2;
- (vi) an ITR2;
- and
- (vii) a SS2;
- operably linked to each other in a 5′-to-3′ direction as:
- SS1-ITR1-HR1-HPM-HR2-ITR2-SS2, wherein the SS1 and the SS2 each, independently, comprise a portion having at least 85% sequence identity to a nucleic acid sequence selected from the group consisting of the nucleic acid sequences of SEQ ID NOS:
- 3-5 and 9-11, and wherein neither the SS1 nor the SS2 comprises an open reading frame that encodes more than 100 amino acids.
8 more elements
A nucleic acid molecule comprising the formula of (SS1-ITR1-CS-ITR2-SS2)n, wherein
- (i) the SS1 is a first spacer;
- (ii) the ITR1 is a first inverted terminal repeat;
- (iii) the CS is a cloning site;
- (iv) the ITR2 is a second inverted terminal repeat;
- and
- (v) the SS2 is a second spacer;
- wherein said SS1, said ITR1, said CS, said ITR2, and said SS2 are operably linked to each other in a 5′-to-3′ direction, and wherein the n is an integer greater than 1, wherein the SS1 and the SS2 together comprise a portion that has at least 85% sequence identity to a nucleic acid sequence selected from the group consisting of the nucleic acid sequences of SEQ ID NOS:
- 3-5 and 9-11, and wherein neither the SS1 nor the SS2 comprises an open reading frame that encodes more than 100 amino acids.
5 more elements
A nucleic acid molecule comprising the formula of (SS1-ITR1-HPM-ITR2-SS2)n, wherein
- (i) the SS1 is a first spacer;
- (ii) the ITR1 is a first inverted terminal repeat;
- (iii) the HPM is a heterologous polynucleotide molecule;
- (iv) the ITR2 is a second inverted terminal repeat;
- and
- (v) the SS2 is a second spacer;
- wherein said SS1, said ITR1, said HPM, said ITR2, and said SS2 are operably linked to each other in a 5′-to-3′ direction, and wherein the n is an integer greater than 1, wherein the SS1 and the SS2 together comprise a portion that has at least 85% sequence identity to a nucleic acid sequence selected from the group consisting of the nucleic acid sequences of SEQ ID NOS:
- 3-5 and 9-11, and wherein nether the SS1 nor the SS2 comprises an open reading frame that encodes more than 100 amino acids.
5 more elements
Dependencies are inferred from the claim text.