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In vivo assembly ofDNA-fragments in themoss, Physcomitrellapatens Brian Christopher KingKonstantinos VavitsasNur Kusaira Khairul Ikram2016 год

In vivo assembly of DNA-fragments in the moss, Physcomitrella patens
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Аннотация: Abstract Direct assembly of multiple linear DNA fragments via homologous recombination, a phenomenon known as in vivo assembly or transformation associated recombination, is used in biotechnology to assemble DNA constructs ranging in size from a few kilobases to full synthetic microbial genomes. It has also enabled the complete replacement of eukaryotic chromosomes with heterologous DNA. The moss Physcomitrella patens , a non-vascular and spore producing land plant (Bryophyte), has a well-established capacity for homologous recombination. Here, we demonstrate the in vivo assembly of multiple DNA fragments in P. patens with three examples of effective genome editing: we (i) efficiently deleted a genomic locus for diterpenoid metabolism yielding a biosynthetic knockout, (ii) introduced a salt inducible promoter, and (iii) re-routed endogenous metabolism into the formation of amorphadiene, a precursor of high-value therapeutics. These proof-of-principle experiments pave the way for more complex and increasingly flexible approaches for large-scale metabolic engineering in plant biotechnology.
Год издания: 2016
Авторы: Brian Christopher King, Konstantinos Vavitsas, Nur Kusaira Khairul Ikram, Josephine Schrøder, Lars B. Scharff, Jean-Étienne Bassard, Björn Hamberger, Poul Erik Jensen, Henrik Toft Simonsen
Издательство: Nature Portfolio
Источник: Scientific Reports
Ключевые слова: Plant biochemistry and biosynthesis, Photosynthetic Processes and Mechanisms, Plant tissue culture and regeneration
Другие ссылки: Scientific Reports (PDF)
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univOAK (4 institutions : Université de Strasbourg, Université de Haute Alsace, INSA Strasbourg, Bibliothèque Nationale et Universitaire de Strasbourg) (PDF)
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Technical University of Denmark, DTU Orbit (Technical University of Denmark, DTU) (HTML)
Research at the University of Copenhagen (University of Copenhagen) (HTML)
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