In vivo assembly of DNA-fragments in the moss, Physcomitrella patens

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Dokumenter

  • Brian Christopher King
  • Konstantinos Vavitsas
  • Nur Kusaira Binti Khairul Ikram
  • Josephine Schrøder
  • Scharff, Lars
  • Björn Robert Hamberger
  • Jensen, Poul Erik
  • Henrik Toft Simonsen

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.

OriginalsprogEngelsk
Artikelnummer25030
TidsskriftScientific Reports
Vol/bind6
Antal sider7
ISSN2045-2322
DOI
StatusUdgivet - 2016

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