IMPORTATION OF MITOCHODRIAL PROTEIN BY AN ENHANCED ALLOTOPIC APPROACH

摘要

An expression vector containing appropriate mitochondrion-targeting sequences (MTS) and appropriate 3′UTR sequences provides efficient and stable delivery of a mRNA encoding a protein (CDS) to the mitochondrion of a mammalian cell. The MTS and 3′UTR sequences guide the CDS mRNA from the nuclear compartment of the cell to mitochondrion-bound polysomes, where the CDS is translated. This provides an efficient translocation of a mature functional protein into the mitochondria. A method of targeting mRNA expressed in the nuclear compartment of a mammalian cell to the mitochondrion is also provided. The vector and methods can be used to treat defects in mitochondrial function.

主权项

1. A method of targeting mRNA expressed in the nuclear compartment of a mammalian cell to mitochondrion-bound polysomes said mammalian cell, the method comprising transfecting said mammalian cell with an expression vector,
wherein said vector comprises: (i) a mitochondrion-targeting nucleic acid sequence (MTS); (ii) a nucleic acid sequence encoding said protein in accordance with a universal genetic code (CDS); and (iii) a 3′UTR nucleic acid sequence, located 3′ of said CDS, wherein, said MIS comprises a cDNA sequence of a MIS of a nuclear encoded mitochondrion-targeted mRNA, said 3 UTR nucleic acid sequence comprises a 3′UTR sequence of a nuclear-encoded mitochondrion-targeted mRNA, said vector does not comprise any sequence identical to a 3′UTR of a naturally occurring mRNA which is a nuclear transcribed but not mitochondrion-targeted mRNA, a cDNA sequence of said 3′UTR of a naturally occurring mRNA, or a DNA sequence coding for said 3′UTR of a naturally-occurring mRNA in accordance with the universal genetic code, and said MTS and said 3′ UTR nucleic acid sequence are configured to target mRNA expressed by said expression vector in the nuclear compartment to the mitochondrion-bound polysomes of said mammalian cell.