2016
DOI: 10.1186/s13007-016-0148-0
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Abstract: BackgroundCRISPR-Cas is a recent and powerful addition to the molecular toolbox which allows programmable genome editing. It has been used to modify genes in a wide variety of organisms, but only two alga to date. Here we present a methodology to edit the genome of Thalassiosira pseudonana, a model centric diatom with both ecological significance and high biotechnological potential, using CRISPR-Cas.ResultsA single construct was assembled using Golden Gate cloning. Two sgRNAs were used to introduce a precise 3… Show more

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Cited by 126 publications
(92 citation statements)
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“…Meganucleases (MNs) and transcription activator-like effector nuleases (TALEN) were utilized to induce targeted mutagenesis (TM) of lipid metabolic genes and generated one strain with 45-fold enhancement in triacylglycerol accumulation in P. tricornutum (Daboussi et al, 2014). Recently, Clustered Regularly Interspaced Short Palindromic Repeats and their associated proteins (CRISPR/Cas9) system has been successfully applied in P. tricornutum and T. pseudonana to create stable mutants (Hopes et al, 2016;Nymark et al, 2016), but to our knowledge there was no reports on editing lipid metabolic genes using CRISPR/Cas9 system yet.…”
Section: Genetic Modificationsmentioning
confidence: 99%
“…Meganucleases (MNs) and transcription activator-like effector nuleases (TALEN) were utilized to induce targeted mutagenesis (TM) of lipid metabolic genes and generated one strain with 45-fold enhancement in triacylglycerol accumulation in P. tricornutum (Daboussi et al, 2014). Recently, Clustered Regularly Interspaced Short Palindromic Repeats and their associated proteins (CRISPR/Cas9) system has been successfully applied in P. tricornutum and T. pseudonana to create stable mutants (Hopes et al, 2016;Nymark et al, 2016), but to our knowledge there was no reports on editing lipid metabolic genes using CRISPR/Cas9 system yet.…”
Section: Genetic Modificationsmentioning
confidence: 99%
“…In addition, CRISPR-Cas9 genome editing has recently become feasible in model diatoms [19, 20], and several diatom viruses have been sequenced [21] which may provide useful resources for building tools as so many animal viruses have done before them. Owing to the relative ease of adapting these modern genetic tools to diatoms, several labs are engaging additional tools with promising results, including proximity proteomics, live cell microscopy, and super-resolution fluorescence microscopy [22].…”
Section: Diatoms Are Ready For Their Close-upmentioning
confidence: 99%
“…The development of CRISPR‐Cas based tools for diatoms were aimed towards developing relatively cost effective and easy genome editing tools for studying fundamental diatom biology via reverse genetics as a basis for obtaining improved strains for biotechnology applications . To date, CRISPR‐Cas systems has been reported to be successfully applied in diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana . In P. tricornutum , a codon optimized Cas9 and sgRNA modules were transformed into the host on the same vector without a selectable marker.…”
Section: Diatomsmentioning
confidence: 99%
“…Unlike the electroporation transformation used in Chlamydomonas and Nannochloropsis , biolistic bombardment was used for transformation of diatoms which could fragment the vectors resulting in incomplete introduction of Cas9 gene into the host . Also, co‐transformation of selectable and non‐selectable vectors has resulted in 40% of the transformants to harbor only the plasmid with selectable marker .…”
Section: Diatomsmentioning
confidence: 99%