Recombinant vector and use in gene therapy
OwnerHongzhan Xuinventor
Life of this patent
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Abstract
A recombinant vector for delivering A3G genes into human cells comprising (i) a gene expression block including an A3G gene selected from a wild type A3G gene represented by SEQ ID NO: 1 and a mutant A3G gene and (ii) a group of elements from a modified lentiviral vector including lentiviral regions of packaging signal (ψ, psi), LTRs, RRE, and PBS; wherein said A3G gene is operably linked to the packaging signal (ψ, psi), LTRs, RRE, and PBS.
Claims8
A method for producing a recombinant vector from a vector-producing cell line, wherein the vector comprising mutant Apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (A3G) genes for delivering said mutant A3G genes into human target cells comprising the steps of:
- a) establishing a cell line by culturing a cell line in a culture medium,
- b) transfecting the cell line with supporting plasmids containing a Vif gene, wherein said Vif gene expresses a Vif protein in the cell line,
- c) selecting the cell line stably expressing the Vif protein,
- d) infecting the cell line of step c) with a recombinant vector comprising (i) a gene expression block including a mutant A3G gene and (ii) a group of elements from a modified lentiviral vector including lentiviral regions of packaging signal (ψ, psi), long-term repeats (LTRs), Rev responsive element (RRE), and primer binding site (PBS);
- wherein said mutant A3G gene is operably linked to the packaging signal (ψ, psi), LTRs, RRE, and PBS,
- e) selecting the cell lines stably producing the recombinant vector, and
- f) expressing envelope proteins in the selected cell line of step e) and harvesting the recombinant vector from the culture medium.
Dependencies are inferred from the claim text.