Retroviral vectors carrying senescent cell derived inhibitors 1 (SDI-1)or antisense SDI-1 nucleotide sequences

An isolated producer cell line stably transfected with a retroviral vector encoding an SDI-1 polypeptide or a fragment thereof, said retroviral vector comprising in 5′ to 3′ order:

• (a) a 5′ LTR;
• (b) a nucleic acid sequence encoding an SDI-1 polypeptide or fragment thereof, wherein said SDI-1 polypeptide or fragment thereof inhibits cell proliferation;
• and
• (c) a 3′ LTR region comprising a complete or partial U3 deletion and an insertion in place thereof, wherein said insertion comprises a polylinker sequence into which a regulatory element or a promoter has been cloned, such that said regulatory element or promoter becomes operatively linked to said nucleic acid sequence and regulates expression of said nucleic acid sequence upon infection of a target cell by a recombinant retroviral particle produced by said isolated producer cell line.

A method for producing a recombinant retroviral particle comprising an RNA sequence encoding an SDI-1 polypeptide that inhibits cell proliferation, the method comprising:

• (a) stably transfecting an isolated producer cell line with a retroviral vector comprising in 5′ to 3′ order:
• (1) a 5′ LTR;
• (2) a nucleic acid sequence encoding an SDI-1 polypeptide that inhibits cell proliferation;
• and
• (3) a 3′ LTR region comprising a complete or partial U3 deletion and an insertion in place thereof, wherein said insertion comprises a polylinker sequence into which a regulatory element or a promoter has been cloned;
• and
• (b) producing said recombinant retroviral particle in said stably transfected isolated producer cell line, wherein said retroviral particle comprises said RNA sequence encoding said SDI-1 polypeptide, and further wherein said regulatory element or promoter becomes operatively linked to said nucleic acid sequence and regulates expression of said nucleic acid sequence upon infection of a target cell by said recombinant retroviral particle.

An isolated producer cell line stably transfected with a retroviral vector encoding an SDI-1 polypeptide that inhibits cell proliferation, said retroviral vector comprising in 5′ to 3′ order:

• (a) a 5′ LTR;
• (b) a nucleic acid sequence encoding an SDI-1 polypeptide that inhibits cell proliferation;
• and
• (c) a 3′ LTR region comprising a complete or partial U3 deletion and an insertion in place thereof, wherein said insertion comprises a polylinker sequence into which a regulatory element or a promoter has been cloned, wherein said regulatory element or promoter becomes operatively linked to said nucleic acid sequence and regulates expression of said nucleic acid sequence upon infection of a target cell by a recombinant retroviral particle produced by said isolated producer cell line.

A method for producing a recombinant retroviral particle comprising an RNA sequence encoding amino acids 1 to 71 of human SDI-1, the method comprising:

• (a) stably transfecting an isolated producer cell line with a retroviral vector comprising a DNA sequence encoding amino acids 1–71 of human SDI-1, wherein:
• (i) amino acids 1–71 of human SDI-1 inhibit cell proliferation;
• and
• (ii) said producer cell comprises at least one DNA construct encoding a protein required for retroviral packaging;
• and
• (b) producing said recombinant retroviral particle in said stably transfected isolated producer cell line.

An isolated producer cell line stably transfected with a retroviral vector encoding amino acids 1–71 of human SDI-1, said retroviral vector comprising in 5′ to 3′ order:

• (a) a 5′ LTR;
• (b) a nucleic acid sequence encoding amino acids 1–71 of human SDI-1, wherein said amino acids 1–71 of human SDI-1 inhibit cell proliferation;
• and
• (c) a 3′ LTR region comprising a complete or partial U3 deletion and an insertion in place thereof, wherein said insertion comprises a polylinker sequence into which a regulatory element or a promoter has been cloned, wherein said regulatory element or promoter becomes operatively linked to said nucleic acid sequence and regulates expression of said nucleic acid sequence upon infection of a target cell by a recombinant retroviral particle produced by said isolated producer cell line.

A method for producing a recombinant retroviral particle comprising an RNA sequence encoding amino acids 42 to 58 of human SDI-1, the method comprising:

• (a) stably transfecting an isolated producer cell line with a retroviral vector comprising a DNA sequence encoding amino acids 42–58 of human SDI-1, wherein:
• (i) amino acids 42–58 of human SDI-1 inhibit cell proliferation;
• and
• (ii) said producer cell comprises at least one DNA construct encoding a protein required for retroviral packaging;
• and
• (b) producing said recombinant retroviral particle in said stably transfected isolated producer cell line.

An isolated producer cell line stably transfected with a retroviral vector encoding amino acids 42–58 of human SDI-1, said retroviral vector comprising in 5′ to 3′ order:

• (a) a 5′ LTR;
• (b) a nucleic acid sequence encoding amino acids 42–58 of human SDI-1, wherein said amino acids 42–58 of human SDI-1 inhibit cell proliferation;
• and
• (c) a 3′ LTR region comprising complete or partial U3 deletion and an insertion in place thereof, wherein said insertion comprises a polylinker sequence into which a regulatory element or a promoter has been cloned, wherein said regulatory element or promoter becomes operatively linked to said nucleic acid sequence and regulates expression of said nucleic acid sequence upon infection of a target cell by a recombinant retroviral particle produced by said isolated producer cell line.