TY - JOUR A1 - Eckert, Alexander A1 - Abbasi, Mozhdeh A1 - Mang, Thomas A1 - Saalwächter, Kay A1 - Walther, Andreas T1 - Structure, Mechanical Properties, and Dynamics of Polyethylenoxide/Nanoclay Nacre-Mimetic Nanocomposites JF - Macromolecules N2 - Nacre-mimetic nanocomposites based on high fractions of synthetic high-aspect-ratio nanoclays in combination with polymers are continuously pushing boundaries for advanced material properties, such as high barrier against oxygen, extraordinary mechanical behavior, fire shielding, and glass-like transparency. Additionally, they provide interesting model systems to study polymers under nanoconfinement due to the well-defined layered nanocomposite arrangement. Although the general behavior in terms of forming such layered nanocomposite materials using evaporative self-assembly and controlling the nanoclay gallery spacing by the nanoclay/polymer ratio is understood, some combinations of polymer matrices and nanoclay reinforcement do not comply with the established models. Here, we demonstrate a thorough characterization and analysis of such an unusual polymer/nanoclay pair that falls outside of the general behavior. Poly(ethylene oxide) (PEO) and sodium fluorohectorite form nacre-mimetic, lamellar nanocomposites that are completely transparent and show high mechanical stiffness and high gas barrier, but there is only limited expansion of the nanoclay gallery spacing when adding increasing amounts of polymer. This behavior is maintained for molecular weights of PEO varied over four orders of magnitude and can be traced back to depletion forces. By careful investigation via X-ray diffraction and proton low-resolution solid-state NMR, we are able to quantify the amount of mobile and immobilized polymer species in between the nanoclay galleries and around proposed tactoid stacks embedded in a PEO matrix. We further elucidate the unusual confined polymer dynamics, indicating a relevant role of specific surface interactions. Y1 - 2020 U6 - https://doi.org/10.1021/acs.macromol.9b01931 SN - 1520-5835 VL - 53 IS - 5 SP - 1716 EP - 1725 PB - ACS Publications CY - Washington, DC ER - TY - JOUR A1 - Jildeh, Zaid B. A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef T1 - Sterilization of Objects, Products, and Packaging Surfaces and Their Characterization in Different Fields of Industry: The Status in 2020 JF - physica status solidi (a) applications and materials science N2 - The treatment method to deactivate viable microorganisms from objects or products is termed sterilization. There are multiple forms of sterilization, each intended to be applied for a specific target, which depends on—but not limited to—the thermal, physical, and chemical stability of that target. Herein, an overview on the currently used sterilization processes in the global market is provided. Different sterilization techniques are grouped under a category that describes the method of treatment: radiation (gamma, electron beam, X-ray, and ultraviolet), thermal (dry and moist heat), and chemical (ethylene oxide, ozone, chlorine dioxide, and hydrogen peroxide). For each sterilization process, the typical process parameters as defined by regulations and the mode of antimicrobial activity are summarized. Finally, the recommended microorganisms that are used as biological indicators to validate sterilization processes in accordance with the rules that are established by various regulatory agencies are summarized. KW - bioburdens KW - sterility tests KW - sterilization efficacy KW - sterilization methods KW - validation methods Y1 - 2021 U6 - https://doi.org/10.1002/pssa.202000732 SN - 1862-6319 N1 - Corresponding author: Michael J. Schöning VL - 218 IS - 13 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Molinnus, Denise A1 - Drinic, Aleksander A1 - Iken, Heiko A1 - Kröger, Nadja A1 - Zinser, Max A1 - Smeets, Ralf A1 - Köpf, Marius A1 - Kopp, Alexander A1 - Schöning, Michael Josef T1 - Towards a flexible electrochemical biosensor fabricated from biocompatible Bombyx mori silk JF - Biosensors and Bioelectronics Y1 - 2021 U6 - https://doi.org/10.1016/j.bios.2021.113204 SN - 0956-5663 VL - 183 IS - Art. 113204 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Yoshinobu, Tatsuo A1 - Schöning, Michael Josef T1 - Light-addressable potentiometric sensors (LAPS) for cell monitoring and biosensing JF - Current Opinion in Electrochemistry Y1 - 2021 U6 - https://doi.org/10.1016/j.coelec.2021.100727 SN - 2451-9103 IS - In Press, Journal Pre-proof PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Engemann, Heiko A1 - Du, Shengzhi A1 - Kallweit, Stephan A1 - Cönen, Patrick A1 - Dawar, Harshal T1 - OMNIVIL - an autonomous mobile manipulator for flexible production JF - Sensors Y1 - 2020 SN - 1424-8220 U6 - https://doi.org/10.3390/s20247249 N1 - Special issue: Sensor Networks Applications in Robotics and Mobile Systems VL - 20 IS - 24, art. no. 7249 SP - 1 EP - 30 PB - MDPI CY - Basel ER - TY - JOUR A1 - Maurischat, Andreas T1 - Algebraic independence of the Carlitz period and its hyperderivatives KW - Drinfeld modules KW - t-modules KW - Transcendence KW - Hyperdifferentials Y1 - 2021 N1 - Zweitveröffentlichung. Verlagsveröffentlichung: https://doi.org/10.1016/j.jnt.2022.01.006 SP - 1 EP - 12 ER - TY - JOUR A1 - Wert, Stefan A1 - Iken, Heiko A1 - Schöning, Michael Josef A1 - Matysik, Frank-Michael T1 - Development of a temperature‐pulse enhanced electrochemical glucose biosensor and characterization of its stability via scanning electrochemical microscopy JF - Electroanalysis N2 - Glucose oxidase (GOx) is an enzyme frequently used in glucose biosensors. As increased temperatures can enhance the performance of electrochemical sensors, we investigated the impact of temperature pulses on GOx that was drop-coated on flattened Pt microwires. The wires were heated by an alternating current. The sensitivity towards glucose and the temperature stability of GOx was investigated by amperometry. An up to 22-fold increase of sensitivity was observed. Spatially resolved enzyme activity changes were investigated via scanning electrochemical microscopy. The application of short (<100 ms) heat pulses was associated with less thermal inactivation of the immobilized GOx than long-term heating. Y1 - 2021 U6 - https://doi.org/10.1002/elan.202100089 SN - 1521-4109 IS - Early View PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Lowis, Carsten A1 - Ferguson, Simon A1 - Paulßen, Elisabeth A1 - Hoehr, Cornelia T1 - Improved Sc-44 production in a siphon-style liquid target on a medical cyclotron JF - Applied Radiation and Isotopes Y1 - 2021 U6 - https://doi.org/10.1016/j.apradiso.2021.109675 SN - 0969-8043 VL - 172 IS - Art. 109675 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Waldvogel, Janice A1 - Ritzmann, Ramona A1 - Freyler, Kathrin A1 - Helm, Michael A1 - Monti, Elena A1 - Albracht, Kirsten A1 - Stäudle, Benjamin A1 - Gollhofer, Albert A1 - Narici, Marco T1 - The Anticipation of Gravity in Human Ballistic Movement JF - Frontiers in Physiology N2 - Stretch-shortening type actions are characterized by lengthening of the pre-activated muscle-tendon unit (MTU) in the eccentric phase immediately followed by muscle shortening. Under 1 g, pre-activity before and muscle activity after ground contact, scale muscle stiffness, which is crucial for the recoil properties of the MTU in the subsequent push-off. This study aimed to examine the neuro-mechanical coupling of the stretch-shortening cycle in response to gravity levels ranging from 0.1 to 2 g. During parabolic flights, 17 subjects performed drop jumps while electromyography (EMG) of the lower limb muscles was combined with ultrasound images of the gastrocnemius medialis, 2D kinematics and kinetics to depict changes in energy management and performance. Neuro-mechanical coupling in 1 g was characterized by high magnitudes of pre-activity and eccentric muscle activity allowing an isometric muscle behavior during ground contact. EMG during pre-activity and the concentric phase systematically increased from 0.1 to 1 g. Below 1 g the EMG in the eccentric phase was diminished, leading to muscle lengthening and reduced MTU stretches. Kinetic energy at take-off and performance were decreased compared to 1 g. Above 1 g, reduced EMG in the eccentric phase was accompanied by large MTU and muscle stretch, increased joint flexion amplitudes, energy loss and reduced performance. The energy outcome function established by linear mixed model reveals that the central nervous system regulates the extensor muscles phase- and load-specifically. In conclusion, neuro-mechanical coupling appears to be optimized in 1 g. Below 1 g, the energy outcome is compromised by reduced muscle stiffness. Above 1 g, loading progressively induces muscle lengthening, thus facilitating energy dissipation. Y1 - 2021 U6 - https://doi.org/10.3389/fphys.2021.614060 SN - 1664-042X PB - Frontiers CY - Lausanne ER - TY - JOUR A1 - Bohndick, Carla A1 - Bosse, Elke A1 - Jänsch, Vanessa K. A1 - Barnat, Miriam T1 - How different diversity factors affect the perception of first-year requirements in higher education JF - Frontline Learning Research N2 - In the light of growing university entry rates, higher education institutions not only serve larger numbers of students, but also seek to meet first-year students’ ever more diverse needs. Yet to inform universities how to support the transition to higher education, research only offers limited insights. Current studies tend to either focus on the individual factors that affect student success or they highlight students’ social background and their educational biography in order to examine the achievement of selected, non-traditional groups of students. Both lines of research appear to lack integration and often fail to take organisational diversity into account, such as different types of higher education institutions or degree programmes. For a more comprehensive understanding of student diversity, the present study includes individual, social and organisational factors. To gain insights into their role for the transition to higher education, we examine how the different factors affect the students’ perception of the formal and informal requirements of the first year as more or less difficult to cope with. As the perceived requirements result from both the characteristics of the students and the institutional context, they allow to investigate transition at the interface of the micro and the meso level of higher education. Latent profile analyses revealed that there are no profiles with complex patterns of perception of the first-year requirements, but the identified groups rather differ in the overall level of perceived challenges. Moreover, SEM indicates that the differences in the perception largely depend on the individual factors self-efficacy and volition. Y1 - 2021 U6 - https://doi.org/10.14786/flr.v9i2.667 SN - 2295-3159 VL - 9 IS - 2 SP - 78 EP - 95 PB - EARLI ER - TY - JOUR A1 - Hugenroth, Kristin A1 - Neidlin, Michael A1 - Engelmann, Ulrich M. A1 - Kaufmann, Tim A. S. A1 - Steinseifer, Ulrich A1 - Heilmann, Torsten T1 - Tipless Transseptal Cannula Concept Combines Improved Hemodynamic Properties and Risk‐Reduced Placement: an In Silico Proof‐of‐Concept JF - Artificial Organs Y1 - 2021 U6 - https://doi.org/10.1111/aor.13964 SN - 1525-1594 IS - Accepted Article PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Ayed, Anis Haj A1 - Kusterer, Karsten A1 - Funke, Harald A1 - Keinz, Jan A1 - Bohn, D. T1 - CFD based exploration of the dry-low-NOx hydrogen micromix combustion technology at increased energy densities JF - Propulsion and Power Research KW - Micromix combustion KW - Hydrogen gas turbine KW - Hydrogen combustion KW - High hydrogen combustion KW - Dry-low-NOx (DLN) combustion Y1 - 2017 SN - 2212-540X U6 - https://doi.org/10.1016/j.jppr.2017.01.005 VL - 6 IS - 1 SP - 15 EP - 24 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Morat, Mareike A1 - Faude, Oliver A1 - Hanssen, Henner A1 - Ludyga, Sebastian A1 - Zacher, Jonas A1 - Eibl, Angi A1 - Albracht, Kirsten A1 - Donath, Lars T1 - Agility Training to Integratively Promote Neuromuscular, Cognitive, Cardiovascular and Psychosocial Function in Healthy Older Adults: A Study Protocol of a One-Year Randomized-Controlled Trial JF - International Journal of Environmental Research and Public Health N2 - Exercise training effectively mitigates aging-induced health and fitness impairments. Traditional training recommendations for the elderly focus separately on relevant physiological fitness domains, such as balance, flexibility, strength and endurance. Thus, a more holistic and functional training framework is needed. The proposed agility training concept integratively tackles spatial orientation, stop and go, balance and strength. The presented protocol aims at introducing a two-armed, one-year randomized controlled trial, evaluating the effects of this concept on neuromuscular, cardiovascular, cognitive and psychosocial health outcomes in healthy older adults. Eighty-five participants were enrolled in this ongoing trial. Seventy-nine participants completed baseline testing and were block-randomized to the agility training group or the inactive control group. All participants undergo pre- and post-testing with interim assessment after six months. The intervention group currently receives supervised, group-based agility training twice a week over one year, with progressively demanding perceptual, cognitive and physical exercises. Knee extension strength, reactive balance, dual task gait speed and the Agility Challenge for the Elderly (ACE) serve as primary endpoints and neuromuscular, cognitive, cardiovascular, and psychosocial meassures serve as surrogate secondary outcomes. Our protocol promotes a comprehensive exercise training concept for older adults, that might facilitate stakeholders in health and exercise to stimulate relevant health outcomes without relying on excessively time-consuming physical activity recommendations. KW - agility KW - prevention KW - healthy aging KW - community dwelling KW - psychosocial Y1 - 2020 U6 - https://doi.org/10.3390/ijerph17061853 SN - 1660-4601 VL - 17 IS - 6 SP - 1 EP - 14 PB - MDPI CY - Basel ER - TY - JOUR A1 - Givanoudi, Stella A1 - Cornelis, Peter A1 - Rasschaert, Geertrui A1 - Wackers, Gideon A1 - Iken, Heiko A1 - Rolka, David A1 - Yongabi, Derick A1 - Robbens, Johan A1 - Schöning, Michael Josef A1 - Heyndrickx, Marc A1 - Wagner, Patrick T1 - Selective Campylobacter detection and quantification in poultry: A sensor tool for detecting the cause of a common zoonosis at its source JF - Sensors and Actuators B: Chemical Y1 - 2021 U6 - https://doi.org/10.1016/j.snb.2021.129484 SN - 0925-4005 IS - In Press, Journal Pre-proof SP - Article 129484 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Malinowski, Daniel A1 - Fournier, Yvan A1 - Horbach, Andreas A1 - Frick, Michael A1 - Magliani, Mirko A1 - Kalverkamp, Sebastian A1 - Hildinger, Martin A1 - Spillner, Jan A1 - Behbahani, Mehdi A1 - Hima, Flutura T1 - Computational fluid dynamics analysis of endoluminal aortic perfusion JF - Perfusion N2 - Introduction: In peripheral percutaneous (VA) extracorporeal membrane oxygenation (ECMO) procedures the femoral arteries perfusion route has inherent disadvantages regarding poor upper body perfusion due to watershed. With the advent of new long flexible cannulas an advancement of the tip up to the ascending aorta has become feasible. To investigate the impact of such long endoluminal cannulas on upper body perfusion, a Computational Fluid Dynamics (CFD) study was performed considering different support levels and three cannula positions. Methods: An idealized literature-based- and a real patient proximal aortic geometry including an endoluminal cannula were constructed. The blood flow was considered continuous. Oxygen saturation was set to 80% for the blood coming from the heart and to 100% for the blood leaving the cannula. 50% and 90% venoarterial support levels from the total blood flow rate of 6 l/min were investigated for three different positions of the cannula in the aortic arch. Results: For both geometries, the placement of the cannula in the ascending aorta led to a superior oxygenation of all aortic blood vessels except for the left coronary artery. Cannula placements at the aortic arch and descending aorta could support supra-aortic arteries, but not the coronary arteries. All positions were able to support all branches with saturated blood at 90% flow volume. Conclusions: In accordance with clinical observations CFD analysis reveals, that retrograde advancement of a long endoluminal cannula can considerably improve the oxygenation of the upper body and lead to oxygen saturation distributions similar to those of a central cannulation. KW - computational fluid dynamics analysis KW - simulation KW - endoluminal KW - aortic perfusion KW - extracorporeal membrane oxygenation Y1 - 2022 U6 - https://doi.org/10.1177/02676591221099809 SN - 1477-111X VL - 0 IS - 0 SP - 1 EP - 8 PB - Sage CY - London ER - TY - JOUR A1 - Lenz, Maximilian A1 - Kahmann, Stephanie Lucina A1 - Behbahani, Mehdi A1 - Pennig, Lenhard A1 - Hackl, Michael A1 - Leschinger, Tim A1 - Müller, Lars Peter A1 - Wegmann, Kilian T1 - Influence of rotator cuff preload on fracture configuration in proximal humerus fractures: a proof of concept for fracture simulation JF - Archives of Orthopaedic and Trauma Surgery N2 - Introduction In regard of surgical training, the reproducible simulation of life-like proximal humerus fractures in human cadaveric specimens is desirable. The aim of the present study was to develop a technique that allows simulation of realistic proximal humerus fractures and to analyse the influence of rotator cuff preload on the generated lesions in regards of fracture configuration. Materials and methods Ten cadaveric specimens (6 left, 4 right) were fractured using a custom-made drop-test bench, in two groups. Five specimens were fractured without rotator cuff preload, while the other five were fractured with the tendons of the rotator cuff preloaded with 2 kg each. The humeral shaft and the shortened scapula were potted. The humerus was positioned at 90° of abduction and 10° of internal rotation to simulate a fall on the elevated arm. In two specimens of each group, the emergence of the fractures was documented with high-speed video imaging. Pre-fracture radiographs were taken to evaluate the deltoid-tuberosity index as a measure of bone density. Post-fracture X-rays and CT scans were performed to define the exact fracture configurations. Neer’s classification was used to analyse the fractures. Results In all ten cadaveric specimens life-like proximal humerus fractures were achieved. Two III-part and three IV-part fractures resulted in each group. The preloading of the rotator cuff muscles had no further influence on the fracture configuration. High-speed videos of the fracture simulation revealed identical fracture mechanisms for both groups. We observed a two-step fracture mechanism, with initial impaction of the head segment against the glenoid followed by fracturing of the head and the tuberosities and then with further impaction of the shaft against the acromion, which lead to separation of the tuberosities. Conclusion A high energetic axial impulse can reliably induce realistic proximal humerus fractures in cadaveric specimens. The preload of the rotator cuff muscles had no influence on initial fracture configuration. Therefore, fracture simulation in the proximal humerus is less elaborate. Using the presented technique, pre-fractured specimens are available for real-life surgical education. KW - Proximal humerus fracture KW - Biomechanical simulation KW - Fracture configuration KW - Fracture simulation KW - Rotator cuff Y1 - 2022 U6 - https://doi.org/10.1007/s00402-022-04471-9 SN - 1434-3916 PB - Springer CY - Berlin, Heidelberg ER - TY - JOUR A1 - Chloé, Radermacher A1 - Malyaran, Hanna A1 - Craveiro, Rogerio Bastos A1 - Peglow, Sarah A1 - Behbahani, Mehdi A1 - Pufe, Thomas A1 - Wolf, Michael A1 - Neuss, Sabine T1 - Mechanical loading on cementoblasts: a mini review JF - Osteologie N2 - Orthodontic treatments are concomitant with mechanical forces and thereby cause teeth movements. The applied forces are transmitted to the tooth root and the periodontal ligaments which is compressed on one side and tensed up on the other side. Indeed, strong forces can lead to tooth root resorption and the crown-to-tooth ratio is reduced with the potential for significant clinical impact. The cementum, which covers the tooth root, is a thin mineralized tissue of the periodontium that connects the periodontal ligament with the tooth and is build up by cementoblasts. The impact of tension and compression on these cells is investigated in several in vivo and in vitro studies demonstrating differences in protein expression and signaling pathways. In summary, osteogenic marker changes indicate that cyclic tensile forces support whereas static tension inhibits cementogenesis. Furthermore, cementogenesis experiences the same protein expression changes in static conditions as static tension, but cyclic compression leads to the exact opposite of cyclic tension. Consistent with marker expression changes, the singaling pathways of Wnt/ß-catenin and RANKL/OPG show that tissue compression leads to cementum degradation and tension forces to cementogenesis. However, the cementum, and in particular its cementoblasts, remain a research area which should be explored in more detail to understand the underlying mechanism of bone resorption and remodeling after orthodontic treatments. KW - Cementoblast KW - Compression KW - Tension KW - Mechanotransduction KW - Forces Y1 - 2022 U6 - https://doi.org/10.1055/a-1826-0777 SN - 1019-1291 VL - 31 IS - 2 SP - 111 EP - 118 PB - Thieme CY - Stuttgart ER - TY - JOUR A1 - Emhardt, Selina N. A1 - Jarodzka, Halszka A1 - Brand-Gruwel, Saskia A1 - Drumm, Christian A1 - Niehorster, Diederick C. A1 - van Gog, Tamara T1 - What is my teacher talking about? Effects of displaying the teacher’s gaze and mouse cursor cues in video lectures on students’ learning JF - Journal of Cognitive Psychology N2 - Eye movement modelling examples (EMME) are instructional videos that display a teacher’s eye movements as “gaze cursor” (e.g. a moving dot) superimposed on the learning task. This study investigated if previous findings on the beneficial effects of EMME would extend to online lecture videos and compared the effects of displaying the teacher’s gaze cursor with displaying the more traditional mouse cursor as a tool to guide learners’ attention. Novices (N = 124) studied a pre-recorded video lecture on how to model business processes in a 2 (mouse cursor absent/present) × 2 (gaze cursor absent/present) between-subjects design. Unexpectedly, we did not find significant effects of the presence of gaze or mouse cursors on mental effort and learning. However, participants who watched videos with the gaze cursor found it easier to follow the teacher. Overall, participants responded positively to the gaze cursor, especially when the mouse cursor was not displayed in the video. KW - Instructional design KW - eye movement modelling examples KW - video learning Y1 - 2022 U6 - https://doi.org/10.1080/20445911.2022.2080831 SN - 2044-5911 SP - 1 EP - 19 PB - Routledge, Taylor & Francis Group CY - Abingdon ER - TY - JOUR A1 - Fiedler, Thomas M. A1 - Orzada, Stephan A1 - Flöser, Martina A1 - Rietsch, Stefan H. G. A1 - Schmidt, Simon A1 - Stelter, Jonathan K. A1 - Wittrich, Marco A1 - Quick, Harald H. A1 - Bitz, Andreas A1 - Ladd, Mark E. T1 - Performance and safety assessment of an integrated transmit array for body imaging at 7 T under consideration of specificabsorption rate, tissue temperature, and thermal dose JF - NMR in Biomedicine N2 - In this study, the performance of an integrated body-imaging array for 7 T with 32 radiofrequency (RF) channels under consideration of local specific absorption rate (SAR), tissue temperature, and thermal dose limits was evaluated and the imaging performance was compared with a clinical 3 T body coil. Thirty-two transmit elements were placed in three rings between the bore liner and RF shield of the gradient coil. Slice-selective RF pulse optimizations for B1 shimming and spokes were performed for differently oriented slices in the body under consideration of realistic constraints for power and local SAR. To improve the B1+ homogeneity, safety assessments based on temperature and thermal dose were performed to possibly allow for higher input power for the pulse optimization than permissible with SAR limits. The results showed that using two spokes, the 7 T array outperformed the 3 T birdcage in all the considered regions of interest. However, a significantly higher SAR or lower duty cycle at 7 T is necessary in some cases to achieve similar B1+ homogeneity as at 3 T. The homogeneity in up to 50 cm-long coronal slices can particularly benefit from the high RF shim performance provided by the 32 RF channels. The thermal dose approach increases the allowable input power and the corresponding local SAR, in one example up to 100 W/kg, without limiting the exposure time necessary for an MR examination. In conclusion, the integrated antenna array at 7 T enables a clinical workflow for body imaging and comparable imaging performance to a conventional 3 T clinical body coil. KW - body imaging at 7 T MRI KW - thermal dose KW - tissue temperature KW - transmit antenna arrays Y1 - 2022 U6 - https://doi.org/10.1002/nbm.4656 SN - 0952-3480 (Print) SN - 1099-1492 (Online) VL - 35 IS - 5 SP - 1 EP - 17 PB - Wiley ER - TY - JOUR A1 - Haeger, Gerrit A1 - Grankin, Alina A1 - Wagner, Michaela T1 - Construction of an Aspergillus oryzae triple amylase deletion mutant as a chassis to evaluate industrially relevant amylases using multiplex CRISPR/Cas9 editing technology JF - Applied Research N2 - Aspergillus oryzae is an industrially relevant organism for the secretory production of heterologous enzymes, especially amylases. The activities of potential heterologous amylases, however, cannot be quantified directly from the supernatant due to the high background activity of native α-amylase. This activity is caused by the gene products of amyA, amyB, and amyC. In this study, an in vitro CRISPR/Cas9 system was established in A. oryzae to delete these genes simultaneously. First, pyrG of A. oryzae NSAR1 was mutated by exploiting NHEJ to generate a counter-selection marker. Next, all amylase genes were deleted simultaneously by co-transforming a repair template carrying pyrG of Aspergillus nidulans and flanking sequences of amylase gene loci. The rate of obtained triple knock-outs was 47%. We showed that triple knockouts do not retain any amylase activity in the supernatant. The established in vitro CRISPR/Cas9 system was used to achieve sequence-specific knock-in of target genes. The system was intended to incorporate a single copy of the gene of interest into the desired host for the development of screening methods. Therefore, an integration cassette for the heterologous Fpi amylase was designed to specifically target the amyB locus. The site-specific integration rate of the plasmid was 78%, with exceptional additional integrations. Integration frequency was assessed via qPCR and directly correlated with heterologous amylase activity. Hence, we could compare the efficiency between two different signal peptides. In summary, we present a strategy to exploit CRISPR/Cas9 for gene mutation, multiplex knock-out, and the targeted knock-in of an expression cassette in A. oryzae. Our system provides straightforward strain engineering and paves the way for development of fungal screening systems. KW - aspergillus KW - CRISPR/Cas9 KW - filamentous fungi KW - genome engineering Y1 - 2023 U6 - https://doi.org/10.1002/appl.202200106 SN - 2702-4288 IS - Early View SP - 1 EP - 15 PB - Wiley-VCH ER -