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A Genome-Wide, Mapped Algal Mutant Library Enables High-Throughput Genetic Studies in a Photosynthetic Eukaryote

100 Pages Posted: 10 Apr 2018 Sneak Peek Status: Review Complete

See all articles by Xiaobo Li

Xiaobo Li

Princeton University - Department of Molecular Biology; Stanford University - Department of Plant Biology

Weronika Patena

Princeton University - Department of Molecular Biology; Stanford University - Department of Plant Biology

Friedrich Fauser

Princeton University - Department of Molecular Biology; Stanford University - Department of Plant Biology

Robert E. Jinkerson

Stanford University - Department of Plant Biology; University of California, Riverside (UCR) - Department of Chemical and Environmental Engineering

Shai Saroussi

Stanford University - Department of Plant Biology

Nina Ivanova

Stanford University - Department of Plant Biology

Jacob M. Robertson

Stanford University - Department of Plant Biology; Princeton University - Department of Molecular Biology

Rebecca Yue

Stanford University - Department of Plant Biology

Ru Zhang

Stanford University - Department of Plant Biology; Donald Danforth Plant Science Center

Josep Vilarrasa-Blasi

Stanford University - Department of Plant Biology

Tyler M. Wittkopp

Stanford University - Department of Plant Biology; Stanford University - Department of Biology; Salk Institute for Biological Studies

Silvia Ramundo

University of California, San Francisco (UCSF) - Department of Biochemistry & Biophysics

Sean R. Blum

Stanford University - Department of Plant Biology

Audrey Goh

Princeton University - Department of Molecular Biology

Matthew Laudon

University of Minnesota - St. Paul - Department of Plant and Microbial Biology

Paul A. Lefebvre

University of Minnesota - St. Paul - Department of Plant and Microbial Biology

Arthur R. Grossman

Stanford University - Department of Plant Biology

Martin C. Jonikas

Stanford University - Department of Biology; Carnegie Institution for Science - Department of Plant Biology; Princeton University - Department of Molecular Biology

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Abstract

Photosynthetic organisms provide food and energy for nearly all life on Earth, yet half of their genes remain uncharacterized. Characterization of gene functions could be greatly accelerated by new genetic resources in unicellular model organisms. We have generated a genome-wide, indexed library of mapped insertion mutants for the unicellular alga Chlamydomonas reinhardtii, which includes disruptions in 83% of nuclear genes. The abundance of individual mutants can be tracked with unique DNA barcodes, allowing the library to be screened as a pool. We demonstrated the power of this platform by performing a genome-wide screen that identified 3,109 mutants with defects in photosynthetic growth. Multiple alleles allowed identification of 44 genes required for photosynthesis, 21 of which are novel. Characterization of one of these genes, CPL3, showed it is important for photosystem I activity. The availability of the 62,389 mutants in our library will accelerate characterization of gene functions in photosynthetic eukaryotes.

Suggested Citation

Li, Xiaobo and Patena, Weronika and Fauser, Friedrich and Jinkerson, Robert E. and Saroussi, Shai and Ivanova, Nina and Robertson, Jacob M. and Yue, Rebecca and Zhang, Ru and Vilarrasa-Blasi, Josep and Wittkopp, Tyler M. and Ramundo, Silvia and Blum, Sean R. and Goh, Audrey and Laudon, Matthew and Lefebvre, Paul A. and Grossman, Arthur R. and Jonikas, Martin C., A Genome-Wide, Mapped Algal Mutant Library Enables High-Throughput Genetic Studies in a Photosynthetic Eukaryote (2018). Available at SSRN: https://ssrn.com/abstract=3155767 or http://dx.doi.org/10.2139/ssrn.3155767
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Xiaobo Li

Princeton University - Department of Molecular Biology

119 Lewis Thomas Laboratory
Washington Road
Princeton, NJ 08544-1014
United States

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Weronika Patena

Princeton University - Department of Molecular Biology

119 Lewis Thomas Laboratory
Washington Road
Princeton, NJ 08544-1014
United States

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Friedrich Fauser

Princeton University - Department of Molecular Biology

119 Lewis Thomas Laboratory
Washington Road
Princeton, NJ 08544-1014
United States

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Robert E. Jinkerson

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

University of California, Riverside (UCR) - Department of Chemical and Environmental Engineering

446 Winston Chung Hall
Riverside, CA 92521
United States

Shai Saroussi

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Nina Ivanova

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Jacob M. Robertson

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Princeton University - Department of Molecular Biology

119 Lewis Thomas Laboratory
Washington Road
Princeton, NJ 08544-1014
United States

Rebecca Yue

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Ru Zhang

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Donald Danforth Plant Science Center

975 North Warson Road
St. Louis, MO 63132
United States

Josep Vilarrasa-Blasi

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Tyler M. Wittkopp

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Stanford University - Department of Biology

Gilbert Building, Rm 109
371 Serra Mall
Stanford, CA 94305
United States

Salk Institute for Biological Studies

10010 North Torrey Pines Road
La Jolla, CA 92037
United States

Silvia Ramundo

University of California, San Francisco (UCSF) - Department of Biochemistry & Biophysics

Mission Bay Campus, Genentech Hall
600 16th St
San Francisco, CA 94143
United States

Sean R. Blum

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Audrey Goh

Princeton University - Department of Molecular Biology

119 Lewis Thomas Laboratory
Washington Road
Princeton, NJ 08544-1014
United States

Matthew Laudon

University of Minnesota - St. Paul - Department of Plant and Microbial Biology

140 Gortner Laboratory
1479 Gortner Avenue
St. Paul, MN 55108
United States

Paul A. Lefebvre

University of Minnesota - St. Paul - Department of Plant and Microbial Biology

140 Gortner Laboratory
1479 Gortner Avenue
St. Paul, MN 55108
United States

Arthur R. Grossman

Stanford University - Department of Plant Biology

260 Panama St
Stanford, CA 94305
United States

Martin C. Jonikas (Contact Author)

Stanford University - Department of Biology ( email )

Gilbert Building, Rm 109
371 Serra Mall
Stanford, CA 94305
United States

Carnegie Institution for Science - Department of Plant Biology ( email )

260 Panama St.
Stanford, CA 94305
United States

Princeton University - Department of Molecular Biology ( email )

119 Lewis Thomas Laboratory
Washington Road
Princeton, NJ 08544-1014
United States