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High-efficiencygenomeediting in plants mediated by a Cas9 gene containing multiple introns

Plant Communications 2021年第2期2卷 63-77页

作者： ramona grützner Patrick Martin Claudia Horn Samuel Mortensen Erin J.Cram Carolyn W.T.Lee-Parsons Johannes Stuttmann Sylvestre Marillonnet Department of Cell and Metabolic Biology Leibniz Institute of Plant BiochemistryWeinberg 306120 Halle(Saale)Germany Institute for Biology Department of Plant GeneticsMartin Luther University Halle-WittenbergWeinbergweg 1006120 Halle(Saale)Germany Department of Biology Northeastern UniversityBostonMAUSA Department of Chemistry and Chemical Biology Northeastern UniversityBostonMAUSA Department of Chemical Engineering Northeastern UniversityBostonMAUSA

The recent discovery of the mode of action of the CRISPR/Cas9 systemhas provided biologists with a useful tool for generating site-specific mutations in genes of *** plants,site-targeted mutations are usually obtained by the stable transformation of a Cas9 expression construct into the plant *** efficiency of introducing mutations in genes of interest can vary considerably depending on the specific features of the constructs,including the source and nature of the promoters and terminators used for the expression of the Cas9 gene and the guide RNA,and the sequence of the Cas9 nuclease *** optimize the efficiency of the Cas9 nuclease in generating mutations in target genes in Arabidopsis thaliana,we investigated several features of its nucleotide and/or amino acid sequence,including the codon usage,the number of nuclear localization signals(NLSs),and the presence or absence of *** found that the Cas9 gene codon usage had some effect on its activity and that two NLSs worked better than ***,the highest efficiency of the constructs was achieved by the addition of 13 introns into the Cas9 coding sequence,which dramatically improved the editing efficiency of the *** of the primary transformants obtained with a Cas9 gene lacking introns displayed a knockout mutant phenotype,whereas between 70%and 100%of the primary transformants generated with the intronized Cas9 gene displayed mutant *** intronized Cas9 gene was also found to be effective in other plants such as Nicotiana benthamiana and Catharanthus roseus.

关键词： CRISPR Cas9 targeted mutagenesis gene targeting

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Efficient scar-free knock-ins of several kilobases in plants by engineered CRISPR-Cas endonucleases

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Molecular Plant 2024年第5期17卷 824-837页

作者： Tom Schreiber Anja Prange Petra Schafer Thomas Iwen ramona grutzner Sylvestre Marillonnet Aurelie Lepage Marie Javelle Wyatt Paul Alain Tissier Department of Cell and Metabolic Biology Leibniz Institute of Plant BiochemistryWeinberg 306120 Halle(Saale)Germany Limagrain Centre de RechercheRoute d’EnnezatCS 9012663720 ChappesFrance

In plants and mammals,non-homologous end-joining is the dominant pathway to repair DNA doublestrand breaks,making it challenging to generate knock-in *** this study,we identified two groups of exonucleases from the herpes virus and the bacteriophage T7 families that conferred an up to 38-fold increase in homology-directed repair frequencies when fused to Cas9/Cas12a in a tobacco mosaic virus-based transient assay in Nicotiana *** achieved precise and scar-free insertion of several kilobases of DNA both in transient and stable transformation *** Arabidopsis thaliana,fusion of Cas9 to a herpes virus family exonuclease led to 10-fold higher frequencies of knock-ins in the first generation of *** addition,we demonstrated stable and heritable knock-ins in wheat in 1%of the primary *** together,our results open perspectives for the routine production of heritable knock-in and gene replacement events in plants.

关键词： homology-directed repair knockin gene replacement CRISPR-Cas 50 exonuclease plants

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Lipid peroxidation: Its effects on the formulation and use of pharmaceutical emulsions

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Asian Journal of Pharmaceutical Sciences 2017年第5期12卷 401-411页

作者： ramona Khanum Haema Thevanayagam School of Postgraduate Studies International Medical University (IMU)

Pharmaceutical delivery systems are developed to improve the physicochemical properties of therapeutic compounds. Emulsions are one of these drug delivering systems formulated using water, oils and lipids as main ingredients. Extensive data are usually generated on the physical and chemical characteristics of these oil-in-water and lipid emulsions. However, the oxidative tendency of emulsions is often overlooked. Oxidation impacts the overall quality and safety of these pharmaceutical emulsions. Additionally, introducing oxidatively unstable emulsions into biological systems further promotes oxidation in situ. Products of these reactions then continue to pose serious harm to cells and fuel other physiological oxidation reactions. Consequently, the increase of oxidation products leads to oxidative damage to biological systems. Thus, emulsions with lower lipid peroxidation are more stable and will reduce the negative effects of oxidation in situ. Preventive measures during the formulation of emulsions are important. Many naturally occurring and cost effective substances possess low oxidation tendencies and confer oxidative protection when used in emulsions. Additionally,certain preparatory methods should be employed to reduce or better control lipid ***, emulsions must be evaluated for their oxidation susceptibility using the various techniques available. Careful attention to the preparation of emulsions and assessment of their oxidative stability will help produce safer emulsions without compromising efficacy.

关键词： Drug delivery Emulsions Lipids Peroxidation Oxidative stress

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Intrinsic charm production of doubly charmed baryons:Collider vs. fixed-target

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Science China(Physics,Mechanics & Astronomy) 2020年第2期63卷 29-31页

作者： ramona Vogt Stanley J.Brodsky 无 Nuclear and Chemical Sciences Division Lawrence Livermore National LaboratoryLivermoreCA 94551USA Department of Physics University of CaliforniaDavisCA 95616USA Stanford Linear Accelerator Center Stanford UniversityStanfordCA 94309USA

The LHCb Collaboration has measured the doubly charmed baryon,■^++cc(ucc),through two different decay channels[1,2]in p+p collisions at center-of-mass energies of 7,8 and 13 *** they have performed similar searches for the doubly charmed baryon ■^+cc(dcc),it has not yet been observed by LHCb[3].However,the fixed-target experiment,SELEX,at Fermilab,reported the observation of the ■^+cc in two different decay channels with a 600 GeV charged hyperon beam[4,5].The beam was composed of an admixture of π^±,p,p components as well as the Σ^- *** discussion has arisen over the fact that the fixed-target observations have not been easily reproduced by the collider experiments at higher energies,where one might have expected the production rates to be much higher due to the orders of magnitude increase in the center of mass energy with LHCb.

关键词： doubly fixed decay

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Stretching on chromosomes sheds light on their architecture

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Signal Transduction and Targeted Therapy 2022年第9期7卷 3179-3180页

作者： ramona Juhlen Wolfram Antonin Institute of Biochemistry and Molecular Cell Biology Medical SchoolRWTH Aachen UniversityAachenGermany

In a recent study published in Nature,Meijering et al.1employ elegant optical trapping and manipulation experiments of mitotic chromosomes to gain insights into their biomechanical *** mitosis,chromosomes rearrange in... 详细信息

关键词： shaped trapping insight

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Production of succinic acid in basket and mobile bed bioreactors-Comparative analysis of substrate mass transfer aspects

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Chinese Journal of Chemical Engineering 2016年第4期24卷 513-520页

作者： Anca-Irina Galaction Dan Cascaval ramona-Mihaela Matran Alexandra Tucaliuc "grigore T. Popa" University of Medicine and Pharmacy of Iasi Faculty of Medical Bioengineering Dept. of Biomedical Science M. Kogalniceanu 9-13 700454 Iasi Romania "Gheorghe Asachi" Technical University of Iasi Faculty of Chemical Engineering and Environmental Protection. Dept. of Biochemical Engineering D. Mangeron 73 700050 Iasi Romania

The glucose mass transfer in the biosynthesis of succinic acid with immobilized Actinobacillus succinogenes cells has been comparatively analyzed for a bioreactor with mobile bed vs. a stationary basket bioreactor. The process has been considered to occur under substrate and product inhibitory effects. The results indicated that the bioreactor with mobile bed is more efficient for biocatalyst particles with a diameter over 3 mm, while the basket bioreactor is more efficient for smaller biocatalyst particles and basket bed thickness below 5 mm. The performances of both configurations of immobilized A. succinogenes cell beds were found to be superior to the column packed bed bioreactor.

关键词： Mass transferDiffusionStirred vesselImmobilized cellsBasket bioreactorSuccinic acid

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Colorectal cancer screening

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World Journal of Gastroenterology 2006年第42期12卷 6747-6750页

作者： ramona M McLoughlin Colm A O'Morain Department of Gas-troenterology Adelaide & Meath Hospital and Trinity College Dublin Ireland

Colorectal cancer is a major public health burden worldwide. There is clear-cut evidence that screening will reduce colorectal cancer mortality and the only contentious issue is which screening tool to use. Most evidence points towards screening with fecal occult blood testing. The immunochemical fecal occult blood tests have a higher sensitivity than the guaiac-based tests. Zn addition, their automation and haemoglobin quantification allows a threshold for colonoscopy to be selected that can be accommodated within individual health care systems.

关键词： Screening Colorectal cancer Fecal occult blood testing Colonoscopy

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Economic growth and nutrition transition: an empirical analysis comparing demand elasticities for foods in China and Russia

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Journal of Integrative Agriculture 2015年第6期14卷 1008-1022页

作者： Christine Burggraf Lena Kuhn ZHAO Qi-ran ramona Teuber Thomas Glauben Leibniz Institute of Agricultural Development in Transition Economies (IAMO) College of Economics and Management China Agricultural University

This study provides empirical evidence on the link between economic growth and nutrition transition in two emerging economies, China and Russia. Both countries have experienced rising average incomes, accompanied by an increasing rate of nutrition-related chronic diseases in recent years. Given the regional heterogeneity between these two countries, we analyze the extent to which income growth as a major driver of nutrition transition has a significant effect on the consumption of different food aggregates and how these effects differ between Chinese and Russian consumers. Our results indicate that with increasing household incomes over time the demand for carbohydrates decreases, while the demand for meat and dairy products, as well as fruits increases. This is a development generally known as nutrition transition. Further, we estimate a Quadratic Almost Ideal Demand System（QUAIDS） for nine different food aggregates for China and Russia. Our results indicate that in both countries all food aggregates have positive expenditure elasticities and are thus normal goods. Moreover, our results indicate that in 2008/2009 meat is still a luxury good in China yet a necessity good in Russia. For 2009, the highest own-price elasticities in China are found for non-meat protein sources and dairy products. Within the meat group, beef, poultry and mutton have the highest price elasticities in China. In Russia, the milk and dairy group, together with the vegetable group, is the most price-elastic food group in 2008. In line with the definition of a nutrition transition, our overall results underscore the finding that income growth in China and Russia tends to increase the demand for animal-based products much stronger than, for example, the demand for carbohydrates. Despite being a positive signal for problems of malnutrition in rural China, this trend of increasing meat consumption might further increase the incidence of chronic diseases in urban areas since there is convincing sc

关键词： Russia nutrition dairy expenditure vegetable considerably empirical household income consumed

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Helium Plasma Effects on Polymer Surfaces:from Plasma Parameters and Surface Properties towards Bioengineering Applications

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Chinese Journal of Polymer Science 2024年第8期42卷 1156-1166页

作者： Andrei Vasile Nastuta Maria Butnaru Byron Cheatham ramona Huzum Vasile Tiron Ionut Topala Physics and Biophysics Education Research Laboratory(P&B-EduResLab) Biomedical Science DepartmentFaculty of Medical Bioengineering‘Grigore T.Popa’University of Medicine and Pharmacy IasiM.Kogalniceanu 9-13Iasi 700454Romania Faculty of Medical Bioengineering ‘Grigore T.Popa’University of Medicine and Pharmacy IasiM.Kogalniceanu 9-13Iasi 700454Romania Cytoviva Inc.570 Devall Drive Suite 301Auburn 36832ALUSA Integrated Center of Environmental Science Studies in the North-Eastern Development Region(CERNESIM) Department of Exact and Natural SciencesInstitute of Interdisciplinary Research‘Alexandru Ioan Cuza’University of IasiBlvd.Carol I no.11Iasi 700506Romania Research Center on Advanced Materials and Technologies(RAMTECH) Department of Exact and Natural SciencesInstitute of Interdisciplinary Research‘Alexandru Ioan Cuza’University of IasiBlvd.Carol I no.11Iasi 700506Romania Iasi Plasma Advanced Research Center(IPARC) Faculty of Physics‘Alexandru Ioan Cuza’University of IasiBlvd.Carol I no.11Iasi 700506Romania

Plasma treatment is necessary to optimize the performance of biomaterial *** enhances and regulates the performance of biomaterial su rfaces,creating an effective interface with the human *** treatments have the ability to modify the chemical com position and physical structu re of a surface while leaving its properties *** possess the ability to modify material su rfaces,eliminate contaminants,conduct investigations on cancer therapy,and facilitate wound *** subject of study in question involves the integration of plasma science and technology with biology and *** a helium plasma jet source,applying up to 18 kV,with an average power of 10 W,polymer foils were treated for 60 *** treatment has the ability to alter the chemical composition and physical structure of a su rface while maintaining its *** investigation involved the application of helium plasma at atmospheric pressure to polyamide 6 and polyethylene terephthalate *** inquiry involves monitoring and assessing the plasma source and polymer materials,as well as analyzing the impacts of plasma *** the mean power of the discharge aids in assessing the economic efficacy of the plasma *** discharge in helium at atmospheric pressure has beneficial effects in technology,where it increases the surface free energy of polymer *** biomedicine,it is used to investigate cytotoxicity and cell survival,particularly in direct blood exposure situations that can expedite *** the specific parameters that influence the plasma source in the desired manner for the intended application is of utmost im portance.

关键词： Atmopheric pressure plasmas Plasma-polymer interaction Plasma Medicine Plasma diagnosis Polymer characterization

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Hybrid Additive Manufacturing of Forming Tools

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Automotive Innovation 2023年第3期6卷 311-323页

作者： Hamed Dardaei Joghan ramona Holker‑Jager Anna Komodromos A.Erman Tekkaya Institute of Forming Technology and Lightweight Components(IUL) TU Dortmund UniversityBaroper Stra.e 30344227 DortmundGermany

Additive manufacturing(AM)is widely used in the automotive industry and has been expanded to include aerospace,marine,and *** flexibility and the possibility of manufacturing complex parts in AM motivate the integration of additive manu-facturing with classical forming technologies,which can improve tooling concepts and reduce *** study presents three applications of this ***,the possibility of successful utilization of selective laser melting for manufacturing extrusion tools with complex cooling channels and paths for thermocouples is reported,leading to significantly reduced inner die temperatures during the extrusion ***,sheet lamination is integrated with laser metal deposition(LMD)to manufacture deep-drawing *** results are achieved in reducing the stair step effect,which is the main challenge in sheet lamination,by LMD and following post-processing such as milling,ball burnishing,and laser *** new manufacturing route shows that LMD can economically and efficiently reduce the stair step effect and omit the hardening step from the conventional manufacturing process ***,LMD is used to manufacture a hot stamping punch with improved surface roughness by ball burnishing and near-surface complex cooling *** experimental results show that the manufactured punch has lower temperatures during hot stamping compared with the conventionally manufactured *** study shows the successful integration of AM processes with classical forming processes.

关键词： Additive manufacturing Laser metal deposition Selective laser melting Rapid tooling Forming tool Layer laminated manufacturing

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