ArticleFEMS yeast research2015
A toolbox of endogenous and heterologous nuclear localization sequences for the methylotrophic yeast Pichia pastoris.
Article in FEMS yeast research, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 24 citations in OpenAlex.
- Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions inJournal of fungi (Basel, Switzerland) · 2024Article
- CRISPR/Cas9-based toolkit for rapid marker recycling and combinatorial libraries in Komagataella phaffii.Applied microbiology and biotechnology · 2024Article
- Engineering of Promoters for Gene Expression in Pichia pastoris.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Late-Stage Functionalisation of Polycyclic (Biomolecules · 2021Article
- Evolution and enrichment of CYP5035 in Polyporales: functionality of an understudied P450 family.Applied microbiology and biotechnology · 2021Article
- Valorisation of pectin-rich agro-industrial residues by yeasts: potential and challenges.Applied microbiology and biotechnology · 2020Review
- Orthologous promoters from related methylotrophic yeasts surpass expression of endogenous promoters of Pichia pastoris.AMB Express · 2020Article
- Modulating CRISPR gene drive activity through nucleocytoplasmic localization of Cas9 inFungal biology and biotechnology · 2019Article
- Expanding the CRISPR/Cas9 toolkit for Pichia pastoris with efficient donor integration and alternative resistance markers.Journal of cellular biochemistry · 2018Article
- Tuning CRISPR-Cas9 Gene Drives inG3 (Bethesda, Md.) · 2018Article
- A streamlined cloning workflow minimising the time-to-strain pipeline for Pichia pastoris.Scientific reports · 2017Article
- Recent advances of molecular toolbox construction expand Pichia pastoris in synthetic biology applications.World journal of microbiology & biotechnology · 2017Review
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Authors and funding
3 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Nuclear localization sequences (NLSs) are required for the import of proteins in the nucleus of eukaryotes. However many proteins from bacteria or bacteriophages are used for basic studies in molecular biology, to generate synthetic genetic circuits or for genome editing applications. Prokaryotic recombinases, CRISPR-associated proteins such as Cas9 or bacterial and viral polymerases require efficient NLSs to function in eukaryotes. The yeast Pichia pastoris is a widely used expression platform for heterologous protein production, but molecular tools such as NLSs are limited. Here we have characterized a set of 10 NLSs for P. pastoris, including the first endogenous NLSs (derived from P. pastoris proteins) and commonly used heterologous NLSs. The NLSs were evaluated by fusing them in N- and C-terminal position to an enhanced green fluorescent protein showing pronounced differences in fluorescence levels and nuclear targeting. Thereby we provide a set of different NLSs that can be applied to optimize the nuclear import of heterologous proteins in P. pastoris, paving the way for the establishment of intricate synthetic biology applications.
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