Compositions and methods for inhibiting human immunodeficiency virus infection by down-regulating human cellular genes

An isolated polynucleotide corresponding to a portion of a gene that encodes 2-oxoglutarate dehydrogenase or a complement thereof, wherein said polynucleotide is operably linked to a regulatory sequence, and expression of said polynucleotide in a host cell inhibits infection by HIV, and wherein the polylnucleotide is produced by a method comprising the steps of:

• (a) randomly fragmenting a polynucleotide encoding 2-oxoglutarate dehydrogenase or a complement thereof, to yield polynucleotide fragments;
• (b) transferring the polynucleotide fragments to an expression vector to yield a polynucleotide fragment library, wherein each of the polynucleotide fragments is operatively linked to a protein translation initiation codon, and wherein the expression vector expresses the polynucleotide fragments in a living eukaryotic cell;
• (c) genetically modifying the living eukaryotic cells by introducing the polynucleotide fragment library into the living eukaryotic cells;
• (d) infecting the living eukaryotic cells with HIV;
• (e) isolating or enriching for genetically modified living eukaryotic cells containing a polynucleotide that inhibits infection of the living eukaryotic cells with HIV;
• and
• (f) obtaining the polynucleotide from the genetically modified cells.

An isolated polynucleotide corresponding to a portion of a gene that encodes pyruvate kinase or a complement thereof, wherein sad polynuclcotide is operably linked to a regulatory sequence, and expression of said polynucleotide in a host cell inhibits infection by HIV, and wherein the polynucleotide is produced by a method comprising the steps of:

• (a) randomly fragmenting a polynucleotide encoding pyruvate kinase or a complement thereof to yield polynucleotide fragments;
• (b) transferring the polynucleotide fragments to an expression vector to yield a polynucleotide fragment library, wherein each of the polynucleotide fragments is operatively linked to a protein translation codon, and wherein the expression vector expresses the polynucleotide fragments in a living eukaryotic cell;
• (c) genetically modifying the living eukaryotic cells by introducing the polynucleotide fragment library into the living eukaryotic cells;
• (d) infecting the living eukaryotic cells with HIV;
• (e) isolating or enriching for genetically modified living eukaryotic cells containing a polynucleotide that inhibits infection of the living eukaryotic cells with HIV;
• and
• (f) obtaining the polynucleotide from the genetically modified cells.

An isolated polynucleotide corresponding to a portion of a gene that encodes calnexin or a complement thereof, wherein said polynucleotide is operably linked to a regulatory sequence, and expression of said polynucleotide in a host cell inhibits infection by HIV, and wherein the polynucleotide is produced by a method comprising the steps of:

• (a) randomly fragmenting a polynucleotide encoding calnexin or a complement thereof to yield polynucleotide fragments;
• (b) transferring the polynucleotide fragments to an expression vector to yield a polynucleotide fragment library, wherein each of the polynucleotide fragments is operatively linked to a protein translation initiation codon, and wherein the expression vector expresses the polynucleotide fragments in a living eukaryotic cell;
• (c) genetically modifying the living eukaryotic cells by introducing, the polynucleotide fragment library into the living eukaryotic cells;
• (d) infecting the living eukaryotic cells with HIV;
• (e) isolating or enriching for genetically modified living eukaryotic cells containg a polynucleotide that inhibits infection of the living eukaryotic cells with HIV;
• and
• (f) obtaining the polynucleotide from the genetically modified cells.

An isolated polynucleotide corresponding to a portion of a gene that encodes ADP-ribosylation factor 3 or a complement thereof, wherein said polynucleotide is operably linked to a regulatory sequence, and expression of said polynucleotide in a host cell inhibits infection by HIV, and wherein the polynucleotide is produced by a method comprising the steps of:

• (a) randomly fragmenting a polynucleotide encoding ADP-ribosylation factor 3 or a complement thereof to yield polynucleotide fragments;
• (b) transferring the polynucleotide to an expression vector to yield a polynucleotide fragment library wherein each of the polynucleotide fragments is operatively linked to a protein translation initiation coding and wherein the expression vector expresses the polynucleotide fragments in a living eukaryotic cell;
• (c) genetically modifying the living eukaryotic cells by introducing the polynucleotide fragment library into the living eukaryotic cells;
• (d) infecting the living eukaryotic cells with HIV;
• (e) isolating or enriching for genetically modified living eukaryotic cells containing a polynucleotide that inhibits infection of the living eukaryotic cells with HIV;
• and
• (f) obtaining the polynucleotide from the genetically modified cells.