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2 oyler, weeks, van etten

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2 oyler, weeks, van etten

  1. 1. Algae  at  the  Nexus  of  21st  Century  Food,   Energy  and  Water  Needs?   George  A.  Oyler  MD  PhD   October  17,  2011   Big  Red  is  The  New  Green  
  2. 2. BioEnergy   ENERGY  Agriculture   Environmental   Products   Enhancement   FOOD   WATER  
  3. 3. Why  Algae  for  Biofuels?  Algae  have  great  potenAal  in  biofuels  and  GHG  abatement   • Algae  have  extremely  high  producJvity.   • Algae  photosyntheJc  and  sequester  CO2.   • Algae  do  not  compete  with  food  crops.   • Algae  do  not  require  farm  land.   • Algae  can  remediate  water.   • Use  of  algae  for  wastewater  treatment  offers  huge  value  added  potenJal.   Caveats:     •   Algae  biofuels  are  not  yet  economically  viable.     •   Major  breakthroughs  in  both  engineering  and  biology  are  required.  
  4. 4. World  human  populaJon  vs  Jme  (human  history)  
  5. 5. •  •  Midwest corn production for biofuels has leadHypoxic to record size of Zone Dead Zone this summer.
  6. 6. Hypoxic zones are a world problem Oxygen-depleted coastal zones Persistent Annual Episodic Periodic
  7. 7. Algae can be combined with animal agriculture, AD, andAquaculture - an integrated industry for Nebraska and Europe Algae   producJon   Aquaculture   producJon  
  8. 8. Algae industry can be phased in with high value products Therapeutics
  9. 9. Highlights  of  Algal  Research  at     The  University  of  Nebraska-­‐Lincoln  Presenter:    Don  Weeks  
  10. 10. Transcriptomic analysis of expression of TAG biosynthesis genes under N deprivation
  11. 11. Raman spectra of algae on a gold surface
  12. 12. Raman image of N-depleted algaeNon-­‐starved  algae  (+N)   N-­‐depleted  algae  (-­‐N)   Triglycerides  
  13. 13. TALEN (TAL Effector Nuclease) technology PotenAal  to  knockout  the  acAvity  of  any  gene  in  any  organism   and  the  potenAal  to  replace  that  gene  with  an  improved   version   inal     F k N-­‐Terminal     A   G   G   T   A   C   T   C   G   A   A   T   C   C   T   N N N oN G   C-­‐Term   N Nucle  I   TAL  Domain   TAL a DN Nse   D N   N omainN oma in   NI   NN   NN   NG   NI   HD   NG   HD   NN   NI   NI   NG   HD   HD   NG   NN   N N N N N N N T A G G T A C T C G A A T C C T G N N N N N N N N N N N N N N N N N N N N N C T C C C A T A T G A A G C G T A N N N N N N N N N N N N N N A T C C A T G A G C T T A G G A C N N N N N N N N N N N N N N N N N N N N N G A G G G T A T A C T T C G C A T N N N N N N N HD   NG   NI   NG   NN   NN   NN   NI   NI   HD   NN   HD   NG   NG   HD   NI   N omainN DNmaN  e   D N   o ins TAL  Domain   TAL a C-­‐Term   N Nucle  I   N N N oN A   C   G   G   G   A   T   A   T   C   T   C   T   G   A   C   N-­‐Terminal     ina k Target  Gene  DNA  Sequence   F l     Double  Strand  DNA  Break   N N N N N N N N N N N NN N N N N N G A A T C C T G N N N N N N N N N N NN N N C T C C C A T A T G A A N T A G G T A C T C C N N N N N N N N N N G G C G T A N N N N N N N N T T A G G A A G G G T A T A C T T C G N N N N N A T G A G C C A T N N N N N A T C C N N N N N N N N N Error-­‐prone  NonHomologous  End-­‐Joining  Repair  N N N N N N NN N N N N N N T A G G T A C T C G A A T C C T G N N N N N N N N N N N N N N C T C C C A T A T G A A G C G T A N N N N N N N N N N N N N NN N N N N N NN N N N N N N A T C C A T G A G C T T A G G A C N N N N N N N N N N N N N N G A G G G T A T A C T T C G C A T N N N N N N N N N N N N N N DeleJons,  InserJons,  Frame-­‐shi  MutaJons   =    Gene  Knockout   Proven  in  Yeast   Now    tesAng  in  algae    
  14. 14. Greenhouse  Algal  Growth  Facility   George  Oyler  
  15. 15. UNL’s  University  and  Commercial  Partners  in  Algae   R&D  (ParJal  List,  U.S.  Only)  Johns  Hopkins  University   CAB-­‐COMM  Collaborators         University  of  California,  San  Diego  Iowa  State  University   University  of  Nebraska-­‐Lincoln  University  of  Maryland   Rutgers  University        (UMBC,  UMBI,  UMD)   Scripps  InsJtuJon  of  Oceanography   University  of  California,  Davis  New  Mexico  State  University   Sapphire  Energy  University  of  California  –  Berkeley   Life  Technologies   Life  Cycle  Associates  University  of  California  –  LA  University  of  Delaware   Other  Commercial  Partners   SynapJc  Research   CGC  (Clean  Green),  LLC  Los  Alamos  NaJonal  Laboratory   Prime  BioSoluJons  Sandia  NaJonal  Laboratory   NEW  Nebraska  Start-­‐up   NanoGreen  Technologies,  LLC  
  16. 16. VIRUSES AND ALGAE: THE GOOD AND THE BAD James L. Van Etten University of NebraskaFood  and  Fuel  Conference  –  University  of  Nebraska     Oct.  17  –  18,  2011  
  17. 17. Viruses Can Serve 3 Functions One bad and two good•  1. Pathogens - bad•  2. Enzymes to degrade cell walls for harvesting lipids – good•  3. Provide genetic elements for altering algae – e.g. promoter & enhancer elements - good
  18. 18. PBCV-1 Replication in Chlorella NC64A     (1-3 min) (3-5 min) 100 nm 100 nm (3-5 hr) (6-8 hr) 500 nm 1 µm
  19. 19. Virus Attaching to & Infecting an Algal Cell
  20. 20. + Virus - Virus
  21. 21. An  Algal  Virus  in  Nature                                                                                                          750 nm
  22. 22. Selected Viruses Infecting Unicellular Algae Size Genome Genome Virus Host Principal references (nm) type size (kb) ChaetocerosCnRNAV 31 ssRNA - Y. Shirai (pers. comm.) neogracilis ChaetocerosCsNIV 38 (ss+ds)DNA 6 Nagasaki et al. (2005c) salsugineum ChlorellaPBCV-1 190 dsDNA 330 Van Etten (2003) variabilis MicromonasMpRHAV 120 dsRNA 25.5 Brussaard C et a. (2004) pusilla Mizumoto et al. (2007), Nagasaki HeterocapsaHcRNAV 30 ssRNA 4.4 et al. (2004b, 2005a, 2006), circularisquama Tomaru et al. (2004a)
  23. 23. Host Cell Wall Degradation Using Chlorella- Virus- Enzymes Control Enzyme treated
  24. 24. Viral Elements for Genetically Engineering Algae•  > Viral promoters elements work very well in plants and bacteria – never tested in algae•  > Viral enhancer element works very well in plants – never tested in algae
  25. 25. Summary Viruses Can Serve 3 Functions•  1.    Pathogens    •  2.  Enzymes  to  degrade  cell  walls  for  harvesAng    lipids  •  3.  Provide  geneAc  elements  for  altering  algae  –    e.g.,  promoter  &  enhancer  elements  

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