Method for modulating gene expression by modifying the cpg content
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Abstract
The invention relates to nucleic acid modifications for a directed expression modulation by the targeted insertion or removal of CpG dinucleotides. The invention also relates to modified nucleic acids and expression vectors.
52 Citations
91 Claims
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1-42. -42. (canceled)
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43. A method for targeted modulation of gene expression, comprising:
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(i) modifying a target nucleic acid molecule by changing the number of CpG dinucleotides present in said target nucleic acid molecule to modulate gene expression, wherein said modifying does not alter the amino acid sequence encoded by said target nucleic acid molecule; (ii) coupling the modified target nucleic acid molecule with a transcription control sequence; (iii) cloning the modified target nucleic acid molecule in an expression vector; and (iv) expressing the modified target nucleic acid molecule in an expression system. - View Dependent Claims (44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65)
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- 66. An isolated nucleic acid molecule having a region capable of transcription, codon-optimized relative to an expression system, wherein the number of CpG dinucleotides is increased compared to the codon-optimized sequence without altering the amino acid sequence encoded by said isolated nucleic acid molecule.
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82. An expression system comprising:
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(a) an isolated nucleic acid molecule having a region capable of transcription, codon-optimized in relation to the expression system, and having a modified number of CpG dinucleotides compared to the codon-optimized sequence, operatively coupled to a transcription control sequence, and (b) an expression environment selected from a cell and a cell-free expression environment, wherein (a) can be expressed, wherein expression of said isolated nucleic acid molecule having an increased number of CpG dinucleotides results in increased expression and expression of said nucleic acid molecule having a reduced number of CpG dinucleotides results in reduced expression. - View Dependent Claims (86, 91)
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Specification