Chinese hamster apoptosis-related genes

Abstract

Provided is an isolated polypeptide comprising a Cricetulus griseus sequence capable of mediating apoptosis of a cell, the sequence being selected from a FAIM sequence shown as SEQ ID NO: 1; a FADD sequence shown as SEQ ID NO: 2; a PDCD6 sequence shown as SEQ ID NO: 3; and a Requiem sequence shown as SEQ ID NO: 4.

1 - 33 . (canceled) 
     
     
         34 . A method of modulating apoptosis of a Chinese Hamster Ovary (CHO) cell, the method comprising modulating expression of a cgRequiem polypeptide in the cell, wherein the cgRequiem polypeptide comprises a sequence having at least 99% sequence identity with SEQ ID NO: 4, thereby modulating apoptosis of the CHO cell. 
     
     
         35 . The method according to  claim 34 , wherein expression of the cgRequiem polypeptide is down-regulated and apoptosis is reduced. 
     
     
         36 . The method according to  claim 34 , wherein the cgRequiem polypeptide is encoded by a polynucleotide comprising a sequence having at least 93% sequence identity to SEQ ID NO: 8; or a sequence which is (i) complementary thereto, (ii) capable of hybridising under stringent conditions thereto, or (iii) degenerate thereto as a result of the genetic code. 
     
     
         37 . The method according to  claim 34 , wherein the cgRequiem polypeptide comprises SEQ ID NO: 4. 
     
     
         38 . The method according to  claim 36 , wherein the polynucleotide comprises SEQ ID NO: 8. 
     
     
         39 . The method according to  claim 35 , wherein the CHO cell is genetically engineered to down-regulate expression of the cgRequiem polypeptide. 
     
     
         40 . The method according to  claim 35 , wherein the CHO cell comprises (i) an anti-sense construct directed against cgRequiem, (ii) a non-functional sequence having at least 99% sequence identity with SEQ ID NO: 4, (iii) a double-stranded (ds) RNA corresponding to a polynucleotide encoding cgRequiem, (iv) a single interfering RNA (siRNA) against cgRequiem or (v) a dominant negative mutant of cgRequiem. 
     
     
         41 . The method according to  claim 40 , wherein the siRNA comprises SEQ ID NO: 15, SEQ ID NO: 16, or SEQ ID NO: 48. 
     
     
         42 . The method according to  claim 34 , wherein the CHO cell comprises a plasmid comprising SEQ ID NO: 40. 
     
     
         43 . The method according to  claim 41 , wherein the sequence is transfected, stably integrated, or transformed into the CHO cell. 
     
     
         44 . The method according to  claim 35 , further comprising expressing a recombinant protein in the CHO cell. 
     
     
         45 . The method according to  claim 44 , wherein the recombinant protein is a heterologous protein. 
     
     
         46 . The method according to  claim 44 , wherein the recombinant protein comprises interferon gamma. 
     
     
         47 . The method according to  claim 44 , wherein: (i) viability of the cell is increased; (ii) yield of the recombinant protein is increased; or (iii) glycosylation of the recombinant protein is increased, compared to a method in which expression of the cgRequiem polypeptide in a cell is not down-regulated. 
     
     
         48 . The method according to  claim 47 , wherein the glycosylation is sialylation. 
     
     
         49 . The method according to  claim 48 , wherein the sialylation of the recombinant protein is greater than 2.9 mol sialic acid/mol of recombinant protein. 
     
     
         50 . The method according to  claim 48 , wherein the sialylation of the recombinant protein is greater than about 3.5 mol of sialic acid/mol of recombinant protein.