The Experts below are selected from a list of 16692 Experts worldwide ranked by ideXlab platform
Jeffry D Sander - One of the best experts on this subject based on the ideXlab platform.
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Engineering Designer transcription activator like effector nucleases talens by real or real fast assembly
Current protocols in molecular biology, 2012Co-Authors: Deepak Reyon, Keith J Joung, Cyd Khayter, Maureen R Regan, Jeffry D SanderAbstract:Engineered transcription activator-like effector nucleases (TALENs) are broadly useful tools for performing targeted genome editing in a wide variety of organisms and cell types including plants, zebrafish, C. elegans, rat, human somatic cells, and human pluripotent stem cells. Here we describe a detailed protocol for the serial, hierarchical assembly of TALENs that requires neither PCR nor specialized multi-fragment ligations and that can be implemented by any laboratory. This restriction enzyme and ligation (REAL) protocol can be practiced using a small plasmid library and user-friendly, web-based software that both identifies target sites in sequences of interest and generates printable graphical guides that facilitate assembly of TALENs. All plasmids required to perform the REAL assembly method are publicly available through the Addgene plasmid distribution service (http://www.addgene.org/talEngineering). Alternatively, to decrease the time and effort required, users can practice REAL using a library of plasmids encoding pre-assembled TALE repeats. With the platform of reagents, protocols, and software we describe, researchers can easily engineer multiple TALENs within two weeks or less using standard cloning techniques.
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Engineering Designer nucleases with customized cleavage specificities
Current protocols in molecular biology, 2011Co-Authors: Jeffry D Sander, Morgan L Maeder, Keith J JoungAbstract:Engineered Designer nucleases can be used to efficiently modify genomic sequence in a wide variety of model organisms and cell types. Zinc finger nucleases (ZFNs), consisting of an engineered zinc finger array fused to a non-specific cleavage domain, have been extensively used to modify a broad range of endogenous genes. Here we describe protocols for Engineering ZFNs targeted to specific gene sequences of interest using the Context-Dependent Assembly (CoDA) method.
Keith J Joung - One of the best experts on this subject based on the ideXlab platform.
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Engineering Designer transcription activator like effector nucleases talens by real or real fast assembly
Current protocols in molecular biology, 2012Co-Authors: Deepak Reyon, Keith J Joung, Cyd Khayter, Maureen R Regan, Jeffry D SanderAbstract:Engineered transcription activator-like effector nucleases (TALENs) are broadly useful tools for performing targeted genome editing in a wide variety of organisms and cell types including plants, zebrafish, C. elegans, rat, human somatic cells, and human pluripotent stem cells. Here we describe a detailed protocol for the serial, hierarchical assembly of TALENs that requires neither PCR nor specialized multi-fragment ligations and that can be implemented by any laboratory. This restriction enzyme and ligation (REAL) protocol can be practiced using a small plasmid library and user-friendly, web-based software that both identifies target sites in sequences of interest and generates printable graphical guides that facilitate assembly of TALENs. All plasmids required to perform the REAL assembly method are publicly available through the Addgene plasmid distribution service (http://www.addgene.org/talEngineering). Alternatively, to decrease the time and effort required, users can practice REAL using a library of plasmids encoding pre-assembled TALE repeats. With the platform of reagents, protocols, and software we describe, researchers can easily engineer multiple TALENs within two weeks or less using standard cloning techniques.
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Engineering Designer nucleases with customized cleavage specificities
Current protocols in molecular biology, 2011Co-Authors: Jeffry D Sander, Morgan L Maeder, Keith J JoungAbstract:Engineered Designer nucleases can be used to efficiently modify genomic sequence in a wide variety of model organisms and cell types. Zinc finger nucleases (ZFNs), consisting of an engineered zinc finger array fused to a non-specific cleavage domain, have been extensively used to modify a broad range of endogenous genes. Here we describe protocols for Engineering ZFNs targeted to specific gene sequences of interest using the Context-Dependent Assembly (CoDA) method.
Saeema Ahmed - One of the best experts on this subject based on the ideXlab platform.
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encouraging reuse of design knowledge a method to index knowledge
Design Studies, 2005Co-Authors: Saeema AhmedAbstract:The long-term aim of this research is to develop a method to index design knowledge that is intuitive to an Engineering Designer and therefore encourage the reuse of information. Eighteen interviews were carried out to understand how Designers described the process of designing a particular component or assembly. The analysis led to the development of a method of indexing design knowledge, which has been evaluated in two stages: (1) evaluation of individual taxonomies within the method; and (2) indexing of 92 reports using the method. The research contributes to the development of an ontology for Engineering design.
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encouraging reuse of design knowledge a method to index knowledge
Design Studies, 2005Co-Authors: Saeema AhmedAbstract:The long-term aim of this research is to develop a method to index design knowledge that is intuitive to an Engineering Designer and therefore encourage the reuse of information. Eighteen interviews were carried out to understand how Designers described the process of designing a particular component or assembly. The analysis led to the development of a method of indexing design knowledge, which has been evaluated in two stages: (1) evaluation of individual taxonomies within the method; and (2) indexing of 92 reports using the method. The research contributes to the development of an ontology for Engineering design.
Brett J Savary - One of the best experts on this subject based on the ideXlab platform.
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Engineering Designer glycomodules for boosting recombinant protein secretion in tobacco hairy root culture and studying hydroxyproline o glycosylation process in plants
Plant Biotechnology Journal, 2019Co-Authors: Ningning Zhang, Tristen Wright, Xiaoting Wang, Uddhab Karki, Brett J SavaryAbstract:The key technical bottleneck for exploiting plant hairy root cultures as a robust bioproduction platform for therapeutic proteins has been low protein productivity, particularly low secreted protein yields. To address this, we engineered novel hydroxyproline (Hyp)-O-glycosylated peptides (HypGPs) into tobacco hairy roots to boost the extracellular secretion of fused proteins and to elucidate Hyp-O-glycosylation process of plant cell wall Hyp-rich glycoproteins. HypGPs representing two major types of cell wall glycoproteins were examined: an extensin module consisting of 18 tandem repeats of 'Ser-Hyp-Hyp-Hyp-Hyp' motif or (SP4)18 and an arabinogalactan protein module consisting of 32 tandem repeats of 'Ser-Hyp' motif or (SP)32 . Each module was expressed in tobacco hairy roots as a fusion to the enhanced green fluorescence protein (EGFP). Hairy root cultures engineered with a HypGP module secreted up to 56-fold greater levels of EGFP, compared with an EGFP control lacking any HypGP module, supporting the function of HypGP modules as a molecular carrier in promoting efficient transport of fused proteins into the culture media. The engineered (SP4)18 and (SP)32 modules underwent Hyp-O-glycosylation with arabino-oligosaccharides and arabinogalactan polysaccharides, respectively, which were essential in facilitating secretion of the fused EGFP protein. Distinct non-Hyp-O-glycosylated (SP4)18 -EGFP and (SP)32 -EGFP intermediates were consistently accumulated within the root tissues, indicating a rate-limiting trafficking and/or glycosylation of the engineered HypGP modules. An updated model depicting the intracellular trafficking, Hyp-O-glycosylation and extracellular secretion of extensin-styled (SP4)18 module and AGP-styled (SP)32 module is proposed.
Hakan Petersson - One of the best experts on this subject based on the ideXlab platform.
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template based design analysis an alternative approach for the Engineering Designer to perform computer based design analysis
2016Co-Authors: Hakan PeterssonAbstract:The current trend in industry to encourage Engineering Designers to take an active part in the analysis of their own design solutions is apparent in many companies today, domestically as well as abroad. From a research project with the objective to develop a computer-based design system for the design of lightweight grippers, one of the major difficulties was to overcome the system users’ lack of knowledge and experience in the design of lightweight structures and Computer-Based Design Analysis (CBDA). CBDA here refers to the use of analysis tools such as Finite Element Analysis (FEA) and computer-based structural optimization. In order to handle these difficulties, the author introduced the use of templates. In the given context, a template refers to an especially pre-formatted code, which contains the implemented information/knowledge necessary to perform a specific task on an operational level. It should be noted that the use of templates as a means of support in performing a specific design or analysis task is not a new phenomenon in industrial practice. Inspired by the opportunities provided by the template approach, the main objective set out for the thesis project was to facilitate the active participation of the Engineering Designers in performing CBDA singlehandedly, or in any other organizational setting, by utilizing a Template-Based Design Analysis (TBDA) approach, as an integrated part of their activities within the Engineering design process. The evolutionary research approach for the development of the TBDA approach is based on surveys in Swedish as well as international industry, literature surveys, the development of a Generic Design Analysis (GDA) process model (facilitating integration of the activities between CBDA and Engineering design) and a number of demonstrator projects to deepen the insights into TBDA. Note that as the TBDA approach is intended for use in industrial practice, the approach is independent of specific Engineering design and product development processes utilized in industry. The conclusion of the thesis work clearly supports the claim that TBDA is not only a competitive approach to current alternatives in supporting the Engineering Designers performing CBDA, but also of a complementary nature providing functionality not included in the alternative approaches currently used in industrial practice.
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using templates to support the Engineering Designer performing computer based design analysis
IMECE 2015 International Mechanical Engineering Congress & Exposition November 13-19 2015 Houston Texas USA, 2015Co-Authors: Hakan Petersson, Damien Motte, Robert BjarnemoAbstract:In their quest for a more efficient and effective utilization of the resources allocated to Engineering design projects, and thus to the overall product development project from which the current d ...
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the Engineering Designer in the role of a design analyst an industrial survey
NWC15 – NAFEMS World Congress 2015 San Diego California USA 21-24 June 2015, 2015Co-Authors: Hakan Petersson, Damien Motte, Robert Bjarnemo, Martin ErikssonAbstract:Traditionally, design analysts are solely responsible for all computerbased design analysis (CBDA). CBDA refers to quantitative design analyses utilising computational tools in the Engineering design and development of technical solutions. There are currently limited insights into and knowledge of tools and methods needed to facilitate the use of CBDA by Engineering Designers. In order to gather information on this aspect of CBDA, an industry survey has been performed.77 persons completed the survey (16% affiliated to NAFEMS) open for twelve weeks during October-December, 2014. Around 35% answered that within their companies CBDA is used by Engineering Designers, and 28% of those who are not currently doing so expect to do so in the future. Linear static analysis is the most frequent type of analysis performed by Engineering Designers. The benefits put forward by the respondents in favour of involving Engineering Designers in CBDA are: it allows early evaluation of concept candidates, shortens lead time, frees resources for the analysis department, and reduces costs. 26% of the respondents answered that there is resistance from the analysis department against allowing Engineering Designers to perform CBDA, 19% within the Engineering design de-partment are also against this involvement and 26% answered that there has been no problem associ-ated with this involvement.Even though the Engineering Designer performs CBDA on his/her own, supervision (56%) and quali-ty assurance of the analysis results (59%) is the responsibility of the design analysts. This is also the case regarding the development of tools and methods to be used by the Engineering Designers as well as instruction and training of the Engineering Designers. (Less)