Enhanced productivity by attenuation of chlorophyll binding protein genes

Patent metadata

Field	Value
Publication number	US-10174087-B2
Application number	US-201615299457-A
Country	US
Kind code	B2
Filing date	Oct 20, 2016
Priority date	Oct 20, 2015
Publication date	Jan 8, 2019
Grant date	Jan 8, 2019

Abstract

Mutant photosynthetic algae having increased biomass productivity are provided. The mutants have attenuated expression of violaxanthin chlorophyll a binding proteins (VCP) or fucoxanthin chlorophyll a/c binding proteins (FCP), reduced chlorophyll, higher apparent ETR(II), little to no reduction in Pmax per cell, and decreased NPQ over a wide range of light intensities. Provided herein are constructs for attenuating or disrupting VCP or FCP genes. Also provided are methods of culturing VCP or FCP mutants for the production of biomass or other products.

First claim

We claim:

1. A recombinant or classically-mutagenized Nannochloropsis alga that has attenuated expression of at least one violaxanthin chlorophyll a-binding protein (VCP) gene.

2. The recombinant or classically-mutagenized mutant Nannochloropsis alga of claim 1 , wherein expression of all VCP genes of the alga are attenuated.

3. The recombinant or classically-mutagenized mutant Nannochloropsis alga of claim 1 , wherein the amount of RNA transcribed by all of the VCP genes in the mutant alga is less than 10% of the amount of RNA transcribed by all of the VCP genes in a control alga.

4. A recombinant or classically-mutagenized Nannochloropsis alga according to claim 1 , wherein the alga has at least one disrupted VCP gene.

5. The recombinant or classically-mutagenized Nannochloropsis alga of claim 4 , wherein all VCP genes of the alga are disrupted.

6. The recombinant or classically-mutagenized Nannochloropsis alga of claim 4 , wherein the at least one VCP gene is disrupted by insertional mutagenesis.

7. The recombinant or classically-mutagenized Nannochloropsis alga of claim 4 , wherein the at least one gene is disrupted by deletion of all or a portion of one or more VCP genes.

8. The recombinant or classically-mutagenized Nannochloropsis alga of claim 4 , wherein the at least one gene is disrupted by homologous recombination.

9. The recombinant or classically-mutagenized Nannochloropsis alga of claim 4 , wherein the at least one gene is disrupted by CRISPR RNA-guided nuclease activity.

10. The recombinant or classically-mutagenized Nannochloropsis alga of claim 1 , wherein the mutant alga exhibits a higher Electron Transport Rate (ETR) than a control alga substantially identical to the mutant alga with the exception that the control alga does not have attenuated expression of at least one VCP gene.

11. The recombinant or classically-mutagenized Nannochloropsis alga of claim 1 , wherein the mutant alga exhibits lower Non-Photochemical Quenching (NPQ) induction than a control alga substantially identical to the alga with the exception that the control alga does not have attenuated expression of at least one VCP gene.

12. The recombinant or classically-mutagenized Nannochloropsis alga of claim 1 , wherein the chlorophyll content of the mutant algal is reduced by at least 15% on a per cell basis.

13. The recombinant or classically-mutagenized Nannochloropsis alga of claim 1 , wherein the mutant alga has increased biomass productivity with respect to a control alga that does not have attenuated expression of at least one VCP gene.

Assignees

• Synthetic Genomics Inc

Inventors

• Meuser Jonathan E
• Dipetrillo Christen G
• Mccarren Jay
• Bailey Shaun

Classifications

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N1/12

  Unicellular algae; Culture media therefor (as new plants A01H13/00) · CPC title

• C07K14/405 Primary

  from algae · CPC title

• C12N2800/80

  Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites · CPC title

• C12N15/902

  using homologous recombination · CPC title