Compact promoters for gene expression
AssigneeHunterian Medicine LLC
InventorVinod Jaskula-Ranga
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Abstract
The invention relates generally to compact promoters and their use in expressing genes, e.g., for treating disease.
Claims78
A nucleic acid comprising a compact promoter operably linked to a coding sequence of a large gene, wherein the compact promoter is between 50 and 250 bp, and wherein the coding sequence of the large gene is greater than about 4110 bp.
The nucleic acid of any preceding claim, wherein the compact promoter comprises a nucleic acid sequence selected from SEQ ID NOs:
- 107-255 or the portion of any one of SEQ ID NOs:
- 25-106, 469-476, 559-564, 609-614, 673-678, 681, 692-697, 706-711, 719-724, 729-734, 748-753, 784-789, 904-909, 920-925, 936-1303 or any sequence in FIGS. 3-19 that corresponds to an H1 promoter (e.g., from about nucleotide 20 to about nucleotide 490 as numbered in FIG. 3), or a nucleic acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or at least about 99.5% identity thereto.
The nucleic acid of any preceding claim, wherein the compact promoter comprises an H1 promoter.
The nucleic acid of any preceding claim, wherein the compact promoter does not comprise a viral promoter and/or a synthetic promoter.
The nucleic acid of any preceding claim, wherein the compact promoter does not comprise F5tg83.
The nucleic acid of any preceding claim, wherein the compact promoter comprises at least 95%, 98%, 99%, 99.5% or 100% identity to a naturally-occurring mammalian promoter.
The nucleic acid of any preceding claim, wherein the compact promoter is capable of expressing a luciferase reporter at a higher level than is a HSK thymidine kinase (TK) promoter.
The nucleic acid of any preceding claim, wherein the coding sequence encodes a cystic fibrosis transmembrane conductance regulator (CFTR), ATP7B, ATP7A, AGL, CPS1, or a functional fragment or variant thereof.
The nucleic acid of any preceding claim, wherein the coding sequence encodes a cystic fibrosis transmembrane conductance regulator (CFTR).
The nucleic acid of any preceding claim, wherein the CFTR coding sequence comprises a truncated form of a wild-type CFTR gene.
An expression construct comprising the nucleic acid of any preceding claim.
A nucleic acid comprising a cystic fibrosis transmembrane conductance regulator (CFTR) coding sequence, wherein the CFTR coding sequence is codon-optimized, wherein the CFTR coding sequence comprises one or more of the following features as compared to a wild type CFTR coding sequence:
- (a) fewer unpaired base pairs of mRNA;
- (b) increased codon usage bias;
- (c) decreased GC content;
- (d) fewer CpG dinucleotides;
- (e) increased mRNA secondary structure;
- (f) fewer cryptic splicing sites;
- (g) fewer premature poly(A) sites;
- (h) fewer RNA instability motifs;
- (i) fewer AT-rich elements (ARE);
- (j) fewer repeat sequences (e.g., direct repeat, reverse repeat, and dyad repeat);
- (k) fewer GC peaks;
- and
- (l) fewer cis-acting elements.
A nucleic acid comprising:
- a) a compact bidirectional promoter;
- b) a protein coding gene;
- and
- c) a second gene;
- wherein the compact bidirectional promoter has a size between 50 and 250 bp, comprises at least one regulatory element that provides for transcription of the protein coding gene in one direction and at least one regulatory element that provides for transcription of the second gene in the other direction, wherein the protein coding gene comprises a coding sequence from about 300 bp to about 4110 bp.
The nucleic acid of any one of claims 55-60, wherein the compact bidirectional promoter comprises an H1 promoter.
Dependencies are inferred from the claim text.