Transgenic microalgae with increased production of at least one omega-3 long chain polyunsaturated fatty acid

Abstract

The invention relates to genetically modified organisms with enhanced production of omega-3 long chain polyunsaturated fatty acids.

1 - 44 . (canceled) 
     
     
         45 . A transgenic organism with increased production of at least one omega-3 LC-PUFAs compared to a control organism, wherein the at least one omega-3 LC-PUFAs is selected from EPA and/or DHA, and wherein the organism expresses a nucleic acid encoding a Δ6-desaturase, a Δ6-elongase and a Δ5-desaturase;
 wherein the nucleic acid encoding Δ6-desaturase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 10 or a sequence with at least 75% homology to SEQ ID NO: 10; 
 wherein the nucleic acid encoding Δ6-elongase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 20 or a sequence with at least 75% homology to SEQ ID NO: 20; and 
 wherein the nucleic acid encoding Δ5-desaturase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 22 or a sequence with at least 75% homology to SEQ ID NO: 22. 
 
     
     
         46 . The transgenic organism of  claim 45 , wherein the organism further expresses a nucleic acid encoding a Δ5-elongase and a Δ4-desaturase, wherein the nucleic acid encoding a Δ5-elongase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 2 or a sequence with at least 75% homology to SEQ ID NO: 2; and wherein the nucleic acid encoding a Δ4-desaturase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 18 or a sequence with at least 75% homology to SEQ ID NO: 18; and wherein the omega-3 LC-PUFAs is DHA. 
     
     
         47 . The transgenic organism of  claim 45 , wherein the organism is a plant, preferably a crop plant. 
     
     
         48 . The transgenic organism according to  claim 47 , wherein the increased DHA content as a percentage of total fatty acids is increased by at least 1%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15% or more compared to a control organism. 
     
     
         49 . The transgenic organism of  claim 46 , wherein the nucleic acid sequence encoding a Δ6-desaturase comprises SEQ ID NO: 9 or a sequence with at least 75% homology to SEQ ID NO: 9; and wherein preferably the nucleic acid sequence encoding a Δ6-elongase comprises SEQ ID NO: 19 or a sequence with at least 75% homology to SEQ ID NO: 19; and wherein preferably the nucleic acid sequence encoding a Δ5-desaturase comprises SEQ ID NO: 21 or a sequence with at least 75% homology to SEQ ID NO: 21; and wherein preferably the nucleic acid sequence encoding a Δ5-elongase comprises SEQ ID NO: 1 or a sequence with at least 75% homology to SEQ ID NO: 11; and wherein preferably the nucleic acid sequence encoding a Δ6-elongase comprises SEQ ID NO: 17 or a sequence with at least 75% homology to SEQ ID NO: 17. 
     
     
         50 . A method for producing a transgenic organism with increased omega-3 LC-PUFA content compared to a control organism, wherein the at least one omega-3 LC-PUFAs is selected from EPA and/or DHA, the method comprising transforming an organism with a nucleic acid encoding a Δ6-desaturase, a Δ6-elongase and a Δ5-desaturase;
 wherein the nucleic acid encoding Δ6-desaturase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 10 or a sequence with at least 75% homology to SEQ ID NO: 10; 
 wherein the nucleic acid encoding Δ6-elongase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 20 or a sequence with at least 75% homology to SEQ ID NO: 20; and 
 wherein the nucleic acid encoding Δ5-desaturase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 22 or a sequence with at least 75% homology to SEQ ID NO: 22. 
 
     
     
         51 . The method of  claim 50 , wherein the method further comprises expressing a nucleic acid encoding a Δ5-elongase and a Δ4-desaturase, wherein the nucleic acid encoding a Δ5-elongase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 2 or a sequence with at least 75% homology to SEQ ID NO: 2; and wherein the nucleic encoding a Δ4-desaturase comprises a nucleic acid encoding the polypeptide of SEQ ID NO: 18 or a sequence with at least 75% homology to SEQ ID NO: 18; and wherein the omega-3 LC-PUFAs is DHA. 
     
     
         52 . A method for increasing production of one of more omega-3 LC-PUFA in an organism, the method comprising
 a) cultivating a transgenic organism according to  claim 45  under conditions which allow for the production of one of more omega-3 LC-PUFAs and   b) obtaining said one of more omega-3 LC-PUFA from the transgenic organism, wherein the omega-3 LC-PUFA is DHA or EPA.   
     
     
         53 . An oil isolated from an organism according to  claim 45  or a foodstuff, feedstuff, nutriceutical or cosmetic obtained from an organism according to  claim 45 . 
     
     
         54 . A composition comprising a transgenic organism according to  claim 45 . 
     
     
         55 . A method of treating or preventing a disorder selected from cardiovascular conditions, including atherosclerosis, thrombosis, high blood pressure, myocardial infarction and atherosclerosis, inflammatory conditions, depression, cognitive decline, arthritis, eczema, metabolic syndrome and type II diabetes, the method comprising administering the composition of  claim 54 . 
     
     
         56 . A method for making a feedstuff comprising a) cultivating a transgenic organism comprising a heterologous transgene as defined in  claim 45  under conditions which allow for the production of one of more omega-3 LC-PUFAs and b) obtaining said one of more omega-3 LC-PUFA from the transgenic organism. 
     
     
         57 . An isolated nucleic acid encoding a Δ6-desaturase (Ost809Δ6), wherein the nucleic acid comprises SEQ ID NO: 9 or a functional variant thereof, wherein the functional variant has at least 75% homology to SEQ ID NO: 9. 
     
     
         58 . A vector or a host cell comprising an isolated nucleic according to  claim 57 , wherein preferably the host cell is an algae or higher plant cell. 
     
     
         59 . An isolated nucleic acid encoding a Δ6-elongase, wherein the nucleic acid comprises SEQ ID NO: 19 or a functional variant thereof, wherein the functional variant has at least 75% homology to SEQ ID NO: 19. 
     
     
         60 . A vector or a host cell comprising an isolated nucleic according to  claim 59 , wherein preferably the host cell is an algae or higher plant cell. 
     
     
         61 . An isolated nucleic acid encoding a Δ5-desaturase, wherein the nucleic acid comprises SEQ ID NO: 21 or a functional variant thereof, wherein the functional variant has at least 75% homology to SEQ ID NO: 21. 
     
     
         62 . A vector or a host cell comprising an isolated nucleic according to  claim 61 , wherein preferably the host cell is an algae or higher plant cell. 
     
     
         63 . An isolated nucleic acid encoding a Δ4-desaturase, wherein the nucleic acid comprises SEQ ID NO: 17 or a functional variant thereof, wherein the functional variant has at least 75% homology to SEQ ID NO: 17. 
     
     
         64 . A vector or a host cell comprising an isolated nucleic according to  claim 63 , wherein preferably the host cell is an algae or higher plant cell.