Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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    Advancements in effect-based surface water quality assessment
    Baat, M.L. De; Oost, R. Van der; Lee, G.H. Van der; Wieringa, N. ; Hamers, T. ; Verdonschot, P.F.M. ; Voogt, P. De; Kraak, M.H.S. - \ 2020
    Water Research 183 (2020). - ISSN 0043-1354
    Agriculture - Bioassay battery - Micropollutants - Passive sampling - Wastewater - Water quality monitoring

    Legally-prescribed chemical monitoring is unfit for determining the pollution status of surface waters, and there is a need for improved assessment methods that consider the aggregated risk of all bioavailable micropollutants present in the aquatic environment. Therefore, the present study aimed to advance effect-based water quality assessment by implementing methodological improvements and to gain insight into contamination source-specific bioanalytical responses. Passive sampling of non-polar and polar organic compounds and metals was applied at 14 surface water locations that were characterized by two major anthropogenic contamination sources, agriculture and wastewater treatment plant (WWTP) effluent, as well as reference locations with a low expected impact from micropollutants. Departing from the experience gained in previous studies, a battery of 20 in vivo and in vitro bioassays was composed and subsequently exposed to the passive sampler extracts. Next, the bioanalytical responses were divided by their respective effect-based trigger values to obtain effect-based risk quotients, which were summed per location. These cumulative ecotoxicological risks were lowest for reference locations (4.3–10.9), followed by agriculture locations (11.3–27.2) and the highest for WWTP locations (12.8–47.7), and were mainly driven by polar organic contaminants. The bioanalytical assessment of the joint risks of metals and (non-)polar organic compounds resulted in the successful identification of pollution source-specific ecotoxicological risk profiles: none of the bioassays were significantly associated with reference locations nor with multiple location types, while horticulture locations were significantly characterized by anti-AR and anti-PR activity and cytotoxicity, and WWTP sites by ERα activity and toxicity in the in vivo bioassays. It is concluded that the presently employed advanced effect-based methods can readily be applied in surface water quality assessment and that the integration of chemical- and effect-based monitoring approaches will foster future-proof water quality assessment strategies on the road to a non-toxic environment.

    Guide-free Cas9 from pathogenic Campylobacter jejuni bacteria causes severe damage to DNA
    Saha, Chinmoy ; Mohanraju, Prarthana ; Stubbs, Andrew ; Dugar, Gaurav ; Hoogstrate, Youri ; Kremers, Gert Jan ; Cappellen, Wiggert A. Van; Horst-Kreft, Deborah ; Laffeber, Charlie ; Lebbink, Joyce H.G. ; Bruens, Serena ; Gaskin, Duncan ; Beerens, Dior ; Klunder, Maarten ; Joosten, Rob ; Demmers, Jeroen A.A. ; Gent, Dik Van; Mouton, Johan W. ; Spek, Peter J. Van Der; Oost, John Van Der; Baarlen, Peter Van; Louwen, Rogier - \ 2020
    Science Advances 6 (2020)25. - ISSN 2375-2548

    CRISPR-Cas9 systems are enriched in human pathogenic bacteria and have been linked to cytotoxicity by an unknown mechanism. Here, we show that upon infection of human cells, Campylobacter jejuni secretes its Cas9 (CjeCas9) nuclease into their cytoplasm. Next, a native nuclear localization signal enables CjeCas9 nuclear entry, where it catalyzes metal-dependent nonspecific DNA cleavage leading to cell death. Compared to CjeCas9, native Cas9 of Streptococcus pyogenes (SpyCas9) is more suitable for guide-dependent editing. However, in human cells, native SpyCas9 may still cause some DNA damage, most likely because of its ssDNA cleavage activity. This side effect can be completely prevented by saturation of SpyCas9 with an appropriate guide RNA, which is only partially effective for CjeCas9. We conclude that CjeCas9 plays an active role in attacking human cells rather than in viral defense. Moreover, these unique catalytic features may therefore make CjeCas9 less suitable for genome editing applications.

    CRISPR with a Happy Ending: Non‐Templated DNA Repair for Prokaryotic Genome Engineering
    Finger‐bou, Max ; Orsi, Enrico ; Oost, John Der; Staals, Raymond H.J. - \ 2020
    Biotechnology Journal 15 (2020). - ISSN 1860-6768
    The exploration of microbial metabolism is expected to support the development of a sustainable economy and tackle several problems related to the burdens of human consumption. Microorganisms have the potential to catalyze processes that are currently unavailable, unsustainable and/or inefficient. Their metabolism can be optimized and further expanded using tools like the clustered regularly interspaced short palindromic repeats and their associated proteins (CRISPR‐Cas) systems. These tools have revolutionized the field of biotechnology, as they greatly streamline the genetic engineering of organisms from all domains of life. CRISPR‐Cas and other nucleases mediate double‐strand DNA breaks, which must be repaired to prevent cell death. In prokaryotes, these breaks can be repaired through either homologous recombination, when a DNA repair template is available, or through template‐independent end joining, of which two major pathways are known. These end joining pathways depend on different sets of proteins and mediate DNA repair with different outcomes. Understanding these DNA repair pathways can be advantageous to steer the results of genome engineering experiments. In this review, we discuss different strategies for the genetic engineering of prokaryotes through either non‐homologous end joining (NHEJ) or alternative end joining (AEJ), both of which are independent of exogenous DNA repair templates
    Off-target activity inhibitors for guided endonucleases
    Notebaart, Richard Alexander ; Künne, Tim Andreas ; Brouns, Stan Johan Jozef ; Oost, John Van Der - \ 2020
    Octrooinummer: WO2020065062, gepubliceerd: 2020-04-02.

    Incidence of off-target DNA cleavage when using CRISPR Cas systems for gene modification are lessened or avoided by using oligonucleotides of rational design and which are antisense to target sequence of the guide RNA or other targeting nucleic acid sequence. Whether in vitro or in vivo, a target nucleic acid comprising a targeted sequence is exposed to the CRISPR enzyme and relevant guiding RNA (or ribonucleoprotein complex) with the antisense oligonucleotide. The antisense oligonucleotide is exposed to the target nucleic acid substantially simultaneously, separately or sequentially together with the CRISPR enzyme or ribonucleoprotein complex.

    From Eat to trEat : Engineering the mitochondrial Eat1 enzyme for enhanced ethyl acetate production in Escherichia coli
    Kruis, Aleksander J. ; Bohnenkamp, Anna C. ; Nap, Bram ; Nielsen, Jochem ; Mars, Astrid E. ; Wijffels, Rene H. ; Oost, John Van Der; Kengen, Servé W.M. ; Weusthuis, Ruud A. - \ 2020
    Biotechnology for Biofuels 13 (2020)1. - ISSN 1754-6834
    Alcohol acetyl transferase (AAT) - Eat1 - Escherichia coli - Ethyl acetate - Mitochondria

    Background: Genetic engineering of microorganisms has become a common practice to establish microbial cell factories for a wide range of compounds. Ethyl acetate is an industrial solvent that is used in several applications, mainly as a biodegradable organic solvent with low toxicity. While ethyl acetate is produced by several natural yeast species, the main mechanism of production has remained elusive until the discovery of Eat1 in Wickerhamomyces anomalus. Unlike other yeast alcohol acetyl transferases (AATs), Eat1 is located in the yeast mitochondria, suggesting that the coding sequence contains a mitochondrial pre-sequence. For expression in prokaryotic hosts such as E. coli, expression of heterologous proteins with eukaryotic signal sequences may not be optimal. Results: Unprocessed and synthetically truncated eat1 variants of Kluyveromyces marxianus and Wickerhamomyces anomalus have been compared in vitro regarding enzyme activity and stability. While the specific activity remained unaffected, half-life improved for several truncated variants. The same variants showed better performance regarding ethyl acetate production when expressed in E. coli. Conclusion: By analysing and predicting the N-terminal pre-sequences of different Eat1 proteins and systematically trimming them, the stability of the enzymes in vitro could be improved, leading to an overall improvement of in vivo ethyl acetate production in E. coli. Truncated variants of eat1 could therefore benefit future engineering approaches towards efficient ethyl acetate production.

    Multilevel optimisation of anaerobic ethyl acetate production in engineered Escherichia coli
    Bohnenkamp, Anna C. ; Kruis, Aleksander J. ; Mars, Astrid E. ; Wijffels, Rene H. ; Oost, John Van Der; Kengen, Servé W.M. ; Weusthuis, Ruud A. - \ 2020
    Biotechnology for Biofuels 13 (2020)1. - ISSN 1754-6834
    Alcohol acetyl transferase (AAT) - Anaerobic - Bioreactor - Eat1 - Escherichia coli - Ethyl acetate - Fermentation

    Background: Ethyl acetate is a widely used industrial solvent that is currently produced by chemical conversions from fossil resources. Several yeast species are able to convert sugars to ethyl acetate under aerobic conditions. However, performing ethyl acetate synthesis anaerobically may result in enhanced production efficiency, making the process economically more viable. Results: We engineered an E. coli strain that is able to convert glucose to ethyl acetate as the main fermentation product under anaerobic conditions. The key enzyme of the pathway is an alcohol acetyltransferase (AAT) that catalyses the formation of ethyl acetate from acetyl-CoA and ethanol. To select a suitable AAT, the ethyl acetate-forming capacities of Atf1 from Saccharomyces cerevisiae, Eat1 from Kluyveromyces marxianus and Eat1 from Wickerhamomyces anomalus were compared. Heterologous expression of the AAT-encoding genes under control of the inducible LacI/T7 and XylS/Pm promoters allowed optimisation of their expression levels. Conclusion: Engineering efforts on protein and fermentation level resulted in an E. coli strain that anaerobically produced 42.8 mM (3.8 g/L) ethyl acetate from glucose with an unprecedented efficiency, i.e. 0.48 C-mol/C-mol or 72% of the maximum pathway yield.

    High-Speed Super-Resolution Imaging Using Protein-Assisted DNA-PAINT
    Filius, Mike ; Cui, Tao Ju ; Ananth, Adithya N. ; Docter, Margreet W. ; Hegge, Jorrit W. ; Oost, John van der; Joo, Chirlmin - \ 2020
    Nano Letters 20 (2020)4. - ISSN 1530-6984 - p. 2264 - 2270.
    Ago-PAINT - Argonaute - DNA origami - DNA-PAINT - single-molecule FRET - super-resolution microscopy

    Super-resolution imaging allows for the visualization of cellular structures on a nanoscale level. DNA-PAINT (DNA point accumulation in nanoscale topology) is a super-resolution method that depends on the binding and unbinding of DNA imager strands. The current DNA-PAINT technique suffers from slow acquisition due to the low binding rate of the imager strands. Here we report on a method where imager strands are loaded into a protein, Argonaute (Ago), which allows for faster binding. Ago preorders the DNA imager strand into a helical conformation, allowing for 10 times faster target binding. Using a 2D DNA origami structure, we demonstrate that Ago-assisted DNA-PAINT (Ago-PAINT) can speed up the current DNA-PAINT technique by an order of magnitude, while maintaining the high spatial resolution. We envision this tool to be useful for super-resolution imaging and other techniques that rely on nucleic acid interactions.

    Nieuw monitoringsmeetnet kwelders Ameland-Oost : Jaarrapportage veldwerk 2019
    Puijenbroek, Marinka E.B. van; Sonneveld, Cor - \ 2020
    Den Helder : Wageningen Marine Research (Wageningen Marine Research rapport C022/20) - 26
    Dit rapport is een update van de vorige jaarrapportage (Duin et al. 2018), waaruit de meer algemene teksten deels zijn overgenomen. Op het Waddeneiland Ameland vindt sinds 1988 bodemdaling plaats als gevolg van gaswinning. Vanaf 1993 vindt er langlopende monitoring plaats naar de mogelijke effecten van de bodemdaling op Ameland-Oost. Een van de conclusies van de laatste integrale rapportage was dat de permanent kwadraten niet geheel representatief waren voor de hele kwelder, en daarom is er in 2019 een nieuw monitoringsplan opgesteld. Onderdeel zijn 80 permanente kwadraten (PQ’s) verspreidt over Neerlands Reid en de Hon waarin in 2019 de eerste vegetatieopnames zijn gemaakt en opslibbingsmetingen met de Sedimentatie-Erosiebalk uitgevoerd. Het ‘oude’ monitoringsnetwerk is voor de laatste keer volledig opgenomen in 2018, maar een aantal PQ’s zijn wel meegenomen in het nieuwe monitoringsnetwerk in 2019. Dit is het eerste jaar dat de nieuwe PQ’s gemeten zijn en opslibbingsresultaten kunnen pas berekend worden na de volgende meting in 2020. In de voorliggende rapportage wordt het nieuwe monitoringsplan geïntroduceerd. Daarnaast worden de laatste gegevens van de opslibbing van het ‘oude’ monitoringsnetwerk voor het jaar 2018 gepresenteerd en de opslibbingsresultaten van 2019 van de ‘oude’ PQ’s die in het nieuwe monitoringsnetwerk zijn opgenomen. Gebaseerd op de metingen uitgevoerd in 2019 van het oude en nieuwe meetnet kunnen we de volgende conclusies trekken: - De opslibbing in 2018 – 2019 was binnen de range van normale waarden voor een eilandkwelder en de sedimentatie volgde het normale patroon met de hoogste waarden aan de wadrand van de kwelder en op de oeverwallen. - In het nieuwe monitoringsnetwerk is op plekken waar regressie (22 PQ’s) heeft plaatsgevonden, vooral middenkwelder veranderd in lage kwelder (14 PQ’s). Er vond meer regressie van vegetatie plaats op het Neerlands Reid (18 PQ’s) dan op de Hon (4 PQ’s). - In kwelderdelen waar tussen 1993 en 2014 regressie van de vegetatie naar lage kwelder heeft plaatsgevonden is de bedekking van kale grond hoger dan in kwelderdelen waar lage kwelder al langer stabiel voorkomt. Een toename aan kale grond slibt minder snel op of erodeert en dat kan tot verdere regressie van de vegetatie leiden.
    Medium-throughput in vitro detection of DNA cleavage by CRISPR-Cas12a
    Creutzburg, Sjoerd C.A. ; Swartjes, Thomas ; Oost, John van der - \ 2020
    Methods : a companion to Methods in enzymology 172 (2020). - ISSN 1046-2023 - p. 27 - 31.
    Cleavage - CRISPR - DNA - Fluorescence - Quantification

    Quantifying DNA cleavage by CRISPR-Cas nucleases is usually done by separating the cleaved products from the non-cleaved target by agarose gel electrophoresis. We devised a method that eliminates the quantification from band intensity on agarose gel, and uses a target with a fluorescent dye on the one end and a biotin on the other. Cleavage of the target will separate the dye from the biotin, and cause the dye to stay in solution when streptavidin beads are introduced. All non-cleaved target will be eliminated from solution and no longer contribute to detectable fluorescence. Cleavage will therefore increase the fluorescent signal. A control, which has no streptavidin treatment, is taken along to correct for any errors that might have been introduced by pipetting, inactivation of the fluorescent dye or release of the biotin during several steps of the procedure. With this method we were able to quantify the fraction of active Cas12a in a purification sample and assess the cleavage rate.

    Highly specific enrichment of rare nucleic acid fractions using Thermus thermophilus argonaute with applications in cancer diagnostics
    Song, Jinzhao ; Hegge, Jorrit W. ; Mauk, Michael G. ; Chen, Junman ; Till, Jacob E. ; Bhagwat, Neha ; Azink, Lotte T. ; Peng, Jing ; Sen, Moen ; Mays, Jazmine ; Carpenter, Erica L. ; Oost, John van der; Bau, Haim H. - \ 2020
    Nucleic acids research 48 (2020)4. - ISSN 0305-1048 - p. e19 - e19.

    Detection of disease-associated, cell-free nucleic acids in body fluids enables early diagnostics, genotyping and personalized therapy, but is challenged by the low concentrations of clinically significant nucleic acids and their sequence homology with abundant wild-type nucleic acids. We describe a novel approach, dubbed NAVIGATER, for increasing the fractions of Nucleic Acids of clinical interest Via DNA-Guided Argonaute from Thermus thermophilus (TtAgo). TtAgo cleaves specifically guide-complementary DNA and RNA with single nucleotide precision, greatly increasing the fractions of rare alleles and, enhancing the sensitivity of downstream detection methods such as ddPCR, sequencing, and clamped enzymatic amplification. We demonstrated 60-fold enrichment of the cancer biomarker KRAS G12D and ∼100-fold increased sensitivity of Peptide Nucleic Acid (PNA) and Xenonucleic Acid (XNA) clamp PCR, enabling detection of low-frequency (<0.01%) mutant alleles (∼1 copy) in blood samples of pancreatic cancer patients. NAVIGATER surpasses Cas9-based assays (e.g. DASH, Depletion of Abundant Sequences by Hybridization), identifying more mutation-positive samples when combined with XNA-PCR. Moreover, TtAgo does not require targets to contain any specific protospacer-adjacent motifs (PAM); is a multi-turnover enzyme; cleaves ssDNA, dsDNA and RNA targets in a single assay; and operates at elevated temperatures, providing high selectivity and compatibility with polymerases.

    First structural insights into CRISPR-Cas-guided DNA transposition
    Oost, John van der; Mougiakos, Ioannis - \ 2020
    Cell Research 30 (2020). - ISSN 1001-0602 - p. 193 - 194.
    The transposition mechanism of the Vibrio cholerae Tn6677 transposon (VcTn6677) is based on the unique synergy between a classical transposition machinery (TnsABC-TniQ complex) and a nuclease-deficient type I-F CRISPR-associated complex for antiviral defense (VcCascade). Four independent studies, three of which appeared in Cell Research, recently reported structures of the VcCascade, VcCascade-TniQ, and/or VcCascade-TniQ-dsDNA complexes as well as a TniQ dimer complex, providing the first insights into the assembly and DNA targeting mechanism of a CRISPR-associated transposase complex.
    Good guide, bad guide: spacer sequence-dependent cleavage efficiency of Cas12a
    Creutzburg, S.C.A. ; Wu, Wen ; Mohanraju, P. ; Swartjes, Thomas ; Alkan, F. ; Gorodkin, J. ; Staals, R.H.J. ; Oost, J. van der - \ 2020
    Nucleic acids research 48 (2020)6. - ISSN 1362-4962 - p. 3228 - 3243.
    Genome editing has recently made a revolutionary development with the introduction of the CRISPR–Cas technology. The programmable CRISPR-associated Cas9 and Cas12a nucleases generate specific dsDNA breaks in the genome, after which host DNA-repair mechanisms can be manipulated to implement the desired editing. Despite this spectacular progress, the efficiency of Cas9/Cas12a-based engineering can still be improved. Here, we address the variation in guide-dependent efficiency of Cas12a, and set out to reveal the molecular basis of this phenomenon. We established a sensitive and robust in vivo targeting assay based on loss of a target plasmid encoding the red fluorescent protein (mRFP). Our results suggest that folding of both the precursor guide (pre-crRNA) and the mature guide (crRNA) have a major influence on Cas12a activity. Especially, base pairing of the direct repeat, other than with itself, was found to be detrimental to the activity of Cas12a. Furthermore, we describe different approaches to minimize base-pairing interactions between the direct repeat and the variable part of the guide. We show that design of the 3′ end of the guide, which is not involved in target strand base pairing, may result in substantial improvement of the guide's targeting potential and hence of its genome editing efficiency.
    Evolutionary classification of CRISPR–Cas systems: a burst of class 2 and derived variants
    Makarova, Kira S. ; Wolf, Yuri I. ; Iranzo, Jaime ; Shmakov, Sergey A. ; Alkhnbashi, Omer S. ; Brouns, Stan J.J. ; Charpentier, Emmanuelle ; Cheng, David ; Haft, Daniel H. ; Horvath, Philippe ; Moineau, Sylvain ; Mojica, Francisco J.M. ; Scott, David ; Shah, Shiraz A. ; Siksnys, Virginijus ; Terns, Michael P. ; Venclovas, Česlovas ; White, Malcolm F. ; Yakunin, Alexander F. ; Yan, Winston ; Zhang, Feng ; Garrett, Roger A. ; Backofen, Rolf ; Oost, John van der; Barrangou, Rodolphe ; Koonin, Eugene V. - \ 2020
    Nature Reviews Microbiology 18 (2020). - ISSN 1740-1526 - p. 67 - 83.

    The number and diversity of known CRISPR–Cas systems have substantially increased in recent years. Here, we provide an updated evolutionary classification of CRISPR–Cas systems and cas genes, with an emphasis on the major developments that have occurred since the publication of the latest classification, in 2015. The new classification includes 2 classes, 6 types and 33 subtypes, compared with 5 types and 16 subtypes in 2015. A key development is the ongoing discovery of multiple, novel class 2 CRISPR–Cas systems, which now include 3 types and 17 subtypes. A second major novelty is the discovery of numerous derived CRISPR–Cas variants, often associated with mobile genetic elements that lack the nucleases required for interference. Some of these variants are involved in RNA-guided transposition, whereas others are predicted to perform functions distinct from adaptive immunity that remain to be characterized experimentally. The third highlight is the discovery of numerous families of ancillary CRISPR-linked genes, often implicated in signal transduction. Together, these findings substantially clarify the functional diversity and evolutionary history of CRISPR–Cas.

    Adaptation and application of a two-plasmid inducible CRISPR-Cas9 system in Clostridium beijerinckii
    Diallo, M. ; Hocq, Rémi ; Collas, Florent ; Chartier, Gwladys ; Wasels, François ; Wijaya, Hani Surya ; Werten, Marc W.T. ; Wolbert, Emil J.H. ; Kengen, Servé W.M. ; Oost, John van der; Ferreira, Nicolas Lopes ; López-Contreras, A.M. - \ 2020
    Methods : a companion to Methods in enzymology 172 (2020). - ISSN 1046-2023 - p. 51 - 60.
    Clostridium beijerinckii - CRISPR-Cas9 - Genome editing - Nuclease

    Recent developments in CRISPR technologies have opened new possibilities for improving genome editing tools dedicated to the Clostridium genus. In this study we adapted a two-plasmid tool based on this technology to enable scarless modification of the genome of two reference strains of Clostridium beijerinckii producing an Acetone/Butanol/Ethanol (ABE) or an Isopropanol/Butanol/Ethanol (IBE) mix of solvents. In the NCIMB 8052 ABE-producing strain, inactivation of the SpoIIE sporulation factor encoding gene resulted in sporulation-deficient mutants, and this phenotype was reverted by complementing the mutant strain with a functional spoIIE gene. Furthermore, the fungal cellulase-encoding celA gene was inserted into the C. beijerinckii NCIMB 8052 chromosome, resulting in mutants with endoglucanase activity. A similar two-plasmid approach was next used to edit the genome of the natural IBE-producing strain C. beijerinckii DSM 6423, which has never been genetically engineered before. Firstly, the catB gene conferring thiamphenicol resistance was deleted to make this strain compatible with our dual-plasmid editing system. As a proof of concept, our dual-plasmid system was then used in C. beijerinckii DSM 6423 ΔcatB to remove the endogenous pNF2 plasmid, which led to a sharp increase of transformation efficiencies.

    Food, nutrition and health in the Netherlands
    Gilissen, Luud J.W.J. - \ 2019
    In: Nutritional and Health Aspects of Food in Western Europe Elsevier - ISBN 9780128131725 - p. 85 - 108.
    Dutch cheese - Grass land - Health and life style - Herring - Hotchpot - Onion - Polder - Potato - Spice trade - Verenigde Oost-Indische Compagnie (VOC) - Whaling

    This chapter provides information on farming, horticulture and fishery in the Netherlands, including its history, and its specific traditional and current food products and related health characteristics. A short description is given on the Dutch agricultural landscape as a result of geological events (depositions of sand, river clay and sea clay) and of human activities (reclaiming large land areas-polders-from inland and sea water areas). In addition, the Dutch North Sea gives ample opportunities for fishing of finfish and shellfish. This sea also opened gates for worldwide (food) trade since several centuries, with great impact in the Dutch Cuisine. Farmer’s food products and the Dutch Cuisine have been described from examples of traditional and local foods in terms of origin, way of production and health characteristics. Nutrition-related conditions have been summarized with regard to life-style; measures for their prevention are described according to the Netherlands Voedingscentrum and illustrated in its Wheel of Five. Finally, some future outlooks have been given on selected Dutch food production systems, and nutrition and health issues.

    De invloed van CRISPR-Cas op de land- en tuinbouw
    Oost, John van der - \ 2019
    Preparing for future AKIS in Europe
    OOst, Inge van; Geerling-Eiff, Floor - \ 2019
    EU - 375 p.
    CRISPR-Cas kansrijke veredelingstool: bacterieel immuunsysteem als biotechnologisch gereedschap
    Oost, John van der - \ 2019
    Editor's cut: DNA cleavage by CRISPR RNA-guided nucleases Cas9 and Cas12a
    Swartjes, Thomas ; Staals, R.H.J. ; Oost, J. van der - \ 2019
    Biochemical Society Transactions 48 (2019)1. - ISSN 0300-5127


    Discovered as an adaptive immune system of prokaryotes, CRISPR–Cas provides many promising applications. DNA-cleaving Cas enzymes like Cas9 and Cas12a, are of great interest for genome editing. The specificity of these DNA nucleases is determined by RNA guides, providing great targeting adaptability. Besides this general method of programmable DNA cleavage, these nucleases have different biochemical characteristics, that can be exploited for different applications. Although Cas nucleases are highly promising, some room for improvement remains. New developments and discoveries like base editing, prime editing, and CRISPR-associated transposons might address some of these challenges.
    Droogte in zandgebieden van Zuid-, Midden- en Oost-Nederland : Rapportage fase 1: ontwikkeling van uniforme werkwijze voor analyse van droogte en tussentijdse bevindingen
    Eertwegh, Gé van den; Bartholomeus, Ruud ; Louw, Perry de; Witte, Flip ; Dam, Jos van; Deijl, Dion van; Hoefsloot, Peter ; Clevers, Sharon ; Hendriks, Dimmie ; Huijgevoort, Marjolein van; Hunink, Joachim ; Mulder, Niels ; Pouwels, Janneke ; Wit, Janine de - \ 2019
    Berg en Dal : KnowH2O - 95
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