Ryan Kalb

Synthetic Genomics (United States)

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W. Marcus Lambert

SUNY Downstate Health Sciences University

John Verruto

VIR Biotechnology (United States)

Eric R. Moellering

Synthetic Genomics (United States)

Imad Ajjawi

Synthetic Genomics (United States)

Kristie Francis

Synthetic Genomics (United States)

Tom J Carlson

Synthetic Genomics (United States)

Gregory Hannum

Synthetic Genomics (United States)

Melisa C. Low

Synthetic Genomics (United States)

Katie Konigsfeld

Synthetic Genomics (United States)

Judit Bartalis

Synthetic Genomics (United States)

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Synthetic Genomics (United States)

VIR Biotechnology (United States)

University of California, San Diego

Cornell University

Stanford University

Michigan State University

Massachusetts Institute of Technology

Massachusetts General Hospital

SUNY Downstate Health Sciences University

State University of New York

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Unrestrained markerless trait stacking in Nannochloropsis gaditana through combined genome editing and marker recycling technologies

Proceedings of the National Academy of Sciences (2018)

John Verruto Kristie Francis Yingjun Wang Melisa C. Low Jessica Greiner

Significance Stacking traits in microalgae is limited by a lack of robust genome modification tools and selectable marker availability. This presents a key hurdle in developing strains for renewable products including biofuels. Here, we overcome these limitations by combining inducible Cre recombinase with constitutive Cas9 nuclease expression in the industrial strain, Nannochloropsis gaditana . With this system, we demonstrate marker- and reporter-free recapitulation of an important lipid productivity trait. In addition, we generate a strain harboring seven-gene knockouts within the photosystem antennae encoding genes. The combined use of relatively mature (Cre) and emerging (CAS9) genome modification technologies can thus accelerate the pace of industrial strain development and facilitate basic research into functionally redundant gene families.

Selectable marker

Gene knockout

Cre recombinase

10.1073/pnas.1718193115

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Citations (76)

Lipid production in Nannochloropsis gaditana is doubled by decreasing expression of a single transcriptional regulator

Nature Biotechnology (2017)

Imad Ajjawi John Verruto Moena Aqui Leah Soriaga Jennifer L. Coppersmith

Insertional mutagenesis

Wild type

10.1038/nbt.3865

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Citations (381)