Collagen producing plants and methods of generating and using same

Abstract

A method of producing collagen in a plant and plants producing collagen are provided. The method is effected by expressing in the plant at least one type of a collagen alpha chain in a manner enabling accumulation of the collagen alpha chain in a subcellular compartment devoid of endogenous P4H activity, thereby producing the collagen in the plant.

1 . A method of producing collagen in a plant or an isolated plant cell comprising targeting to a vacuole of the plant or the isolated plant cell the collagen alpha 1 chain as set forth in SEQ ID NO: 3 and an exogenous prolyl-4-hydroxylase (P4H) so as to allow hydroxylation of the collagen alpha 1 chain by said exogenous P4H and not by an endogenous P4H of the plant or isolated plant cell, thereby producing the collagen in the plant. 
     
     
         2 . The method of  claim 1 , further comprising expressing an exogenous polypeptide selected from the group consisting of Lysyl hydroxylase (LH), protease N and protease C. 
     
     
         3 . The method of  claim 1 , wherein said exogenous P4H comprises a mammalian P4H. 
     
     
         4 . The method of  claim 1 , wherein the plant is selected from the group consisting of Tobacco, Maize, Alfalfa, Rice, Potato, Soybean, Tomato, Wheat, Barley, Canola, Carrot and Cotton. 
     
     
         5 . The method of  claim 1 , wherein said exogenous P4H is capable of specifically hydroxylating the Y position of Gly-X-Y triplets of said at least one type of said collagen chain. 
     
     
         6 . The method of  claim 3 , wherein said mammalian P4H comprises a human P4H. 
     
     
         7 . The method of  claim 1 , wherein the plant is subjected to a stress condition. 
     
     
         8 . The method of  claim 7 , wherein said stress condition is selected from the group consisting of drought, salinity, injury, cold and spraying with stress inducing compounds. 
     
     
         9 . The method of  claim 1 , further comprising targeting to a vacuole of the plant or the isolated plant cell the collagen alpha 2 chain as set forth in SEQ ID NO: 6 so as to allow hydroxylation of the collagen alpha 2 chain by said exogenous P4H and not by an endogenous P4H of the plant or isolated plant cell. 
     
     
         10 . A genetically modified plant or isolated plant cell comprising in a vacuole thereof:
 (i) at least one type of a collagen chain; and   (ii) an exogenous P4H.   
     
     
         11 . A method of producing collagen or comprising:
 (a) providing a plant system comprising:   a first genetically modified plant comprising in a vacuole thereof:   (i) the collagen alpha 1 chain as set forth in SEQ ID NO: 3; and   (ii) an exogenous P4H; and   a second genetically modified plant comprising in a vacuole thereof:   (i) the collagen alpha 2 chain as set forth in SEQ ID NO: 6; and   (ii) an exogenous P4H;   (b) crossing said first plant and said second plant; and   (c) selecting progeny expressing said collagen alpha 1 chain and said collagen alpha 2 chain thereby producing collagen.   
     
     
         12 . The method of  claim 11 , wherein said exogenous P4H is capable of specifically hydroxylating the Y position of Gly-X-Y triplets of said collagen alpha 1 chain or collagen alpha 2 chain. 
     
     
         13 . The method of  claim 11 , wherein said exogenous P4H is human P4H. 
     
     
         14 . A method of producing collagen comprising:
 (a) providing the plant system comprising:   a first genetically modified plant comprising in a vacuole thereof:   (i) the collagen alpha 1 chain as set forth in SEQ ID NO: 3; and   (ii) the collagen alpha 2 chain as set forth in SEQ ID NO: 6; and   a second genetically modified plant comprising in a vacuole thereof an exogenous P4H; and   (b) crossing said first plant and said second plant and selecting progeny expressing the collagen alpha 1 chain, the collagen alpha 2 chain and said P4H thereby producing collagen.