Genetic element E3 for enhanced intracellular expression of target protein encoded in RNA therapeutics
The present invention relates to an intracellular expression enhancing genetic element, which enhances the intracellular expression of target protein encoded in RNA therapeutics such as mRNA vaccines and RNA medicines. In more detail, the present invention relates to the RNA molecule which has the genetic element capable of enhancing the intracellular expression of target protein encoded in RNA therapeutics delivered into cells; the plasmid used to synthesize the RNA molecule having the genetic element “E3”; the method for synthesizing RNA molecule having the genetic element “E3” using the plasmid and utilizing the synthesized RNA molecule as RNA therapeutics.
1. An mRNA molecule having a sequence that functionally enhances the intracellular expression of an encoded target protein, comprising
a) a 5′-untranslated region (UTR) sequence,
b) an AUG sequence for translation initiation,
c) an enhancing genetic element RNA sequence as set forth in SEQ ID NO: 2,
d) a target protein-coding sequence placed adjacent to the 3′-end of the said enhancing genetic element RNA sequence as set forth in SEQ ID NO: 2,
e) a 3′-untranslated region (UTR) sequence, and
f) a poly U sequence.
2. The mRNA molecule of claim 1 , the said Poly U sequence comprises 20-200 nucleotides in length.
3. The mRNA molecule of claim 1 , the RNA molecule is a mRNA vaccine or RNA medicine.
4. A plasmid comprising the arrangement in order of a) a 5′ untranslated region (UTR) sequence, b) an ATG sequence, c) an enhancing genetic element DNA sequence set forth in SEQ ID NO: 1, d) a target protein-coding sequence placed adjacent to 3′-end of the said enhancing genetic element DNA sequence set forth in SEQ ID NO: 1, e) a 3′-untranslated region (UTR) sequence, and f) a poly A sequence.
5. The plasmid of claim 4 , the said poly A sequence comprises 20-200 nucleotides in length.
6. A method for RNA molecule synthesis comprising: RNA molecule synthesis comprising:
introducing, to a cell, a plasmid comprising the arrangement in order of a) a 5′-untranslated region (UTR) sequence, b) a translation initiation codon ATG, c) an enhancing genetic element DNA sequence set forth in SEQ ID NO: 1, d) a target protein-coding sequence placed adjacent to 3′-end of the said enhancing genetic element DNA sequence set forth in SEQ ID NO: 1, e) a 3′-untranslated region (UTR) sequence, and f) a poly A sequence, and
incubating the cell for sufficient time to allow for in vitro transcription of a RNA molecule transcribed from the plasmid, thereby synthesizing the RNA molecule.
7. The method of claim 6 , wherein the said poly A sequence comprises 20-200 nucleotides in length.