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Genome Engineering Via Crispr Cas9 System

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Genome Engineering via CRISPR Cas9 System

Genome Engineering via CRISPR Cas9 System Book
Author : Vijai Singh,Pawan K. Dhar
Publisher : Academic Press
Release : 2020-02-18
ISBN : 0128181419
Language : En, Es, Fr & De

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Book Description :

Genome Engineering via CRISPR-Cas9 Systems presents a compilation of chapters from eminent scientists from across the globe who have established expertise in working with CRISPR-Cas9 systems. Currently, targeted genome engineering is a key technology for basic science, biomedical and industrial applications due to the relative simplicity to which they can be designed, used and applied. However, it is not easy to find relevant information gathered in a single source. The book contains a wide range of applications of CRISPR in research of bacteria, virus, algae, plant and mammalian and also discusses the modeling of drosophila, zebra fish and protozoan, among others. Other topics covered include diagnosis, sensor and therapeutic applications, as well as ethical and regulatory issues. This book is a valuable source not only for beginners in genome engineering, but also researchers, clinicians, stakeholders, policy makers, and practitioners interested in the potential of CRISPR-Cas9 in several fields. Provides basic understanding and a clear picture on how to design, use and implement the CRISPR-Cas9 system in different organisms Explains how to create an animal model for disease research and screening purposes using CRISPR Discusses the application of CRISPR-Cas9 systems in basic sciences, biomedicine, virology, bacteriology, molecular biology, neurology, cancer, industry, and many more

Molecular Aspects of Plant Beneficial Microbes in Agriculture

Molecular Aspects of Plant Beneficial Microbes in Agriculture Book
Author : Vivek Sharma,Richa Salwan
Publisher : Academic Press
Release : 2020-03-12
ISBN : 0128184698
Language : En, Es, Fr & De

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Book Description :

Molecular Aspects of Plant Beneficial Microbes in Agriculture explores their diverse interactions, including the pathogenic and symbiotic relationship which leads to either a decrease or increase in crop productivity. Focusing on these environmentally-friendly approaches, the book explores their potential in changing climatic conditions. It presents the exploration and regulation of beneficial microbes in offering sustainable and alternative solutions to the use of chemicals in agriculture. The beneficial microbes presented here are capable of contributing to nutrient balance, growth regulators, suppressing pathogens, orchestrating immune response and improving crop performance. The book also offers insights into the advancements in DNA technology and bioinformatic approaches which have provided in-depth knowledge about the molecular arsenal involved in mineral uptake, nitrogen fixation, growth promotion and biocontrol attributes.

Plant Genome Editing Policies and Governance

Plant Genome Editing     Policies and Governance Book
Author : Thorben Sprink,Ralf Alexander Wilhelm,Armin Spök,Jürgen Robienski,Stephan Schleissing,Joachim Hermann Schiemann
Publisher : Frontiers Media SA
Release : 2020-04-22
ISBN : 2889636704
Language : En, Es, Fr & De

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The CRISPR Cas9 System for Crop Improvement Progress and Prospects

The CRISPR Cas9 System for Crop Improvement  Progress and Prospects Book
Author : Kah-Yung Bernard Leong
Publisher :
Release : 2018
ISBN :
Language : En, Es, Fr & De

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Book Description :

The global demand for high-quality crops is continuously growing with time. Crop improvement techniques have a long history and they had been applied since the beginning of domestication of the first agricultural plants. Since then, various new techniques have and are being developed to further increase the commercial value and yield of crops. The latest crop improvement technique known as genome editing is a technique that enables precise modification of the plant genome via knocking out undesirable genes or enabling genes to gain new function. The variants generated from the genome editing are indistinguishable from naturally occurring variation. It is also less time-consuming and more readily accepted in the market commercially. The usage of genome editing has proven to be advantages and plays a promising role in future crop improvement efforts. Therefore, in this chapter, we aim to highlight the progress and application of genome editing techniques, in particular, the CRISPR/Cas9 system as a powerful genome editing tool for crop improvement. In addition, the challenges and future prospects of this technology for crop improvement will also be discussed.

SCHWARTZ S PRINCIPLES OF SURGERY 2 volume set 11th edition

SCHWARTZ S PRINCIPLES OF SURGERY 2 volume set 11th edition Book
Author : F. Charles Brunicardi,Dana K. Andersen,Timothy R. Billiar,David L. Dunn,John G. Hunter,Lillian Kao,Jeffrey B. Matthews,Raphael E. Pollock
Publisher : McGraw Hill Professional
Release : 2019-05-29
ISBN : 1259835375
Language : En, Es, Fr & De

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Book Description :

Publisher's Note: Products purchased from Third Party sellers are not guaranteed by the publisher for quality, authenticity, or access to any online entitlements included with the product. The BEST EDITION yet of the #1 text for surgical practice and education For half-a-century, no other text has provided such a solid grounding in basic science, anatomy, operative techniques, and more recently, professional development and leadership training, as Schwartz’s Principles of Surgery. Written by the world’s foremost surgeons, this landmark reference offers distinctly modern and all-encompassing coverage of every important topic in general surgery. Enhanced by a new two volume presentation, the Eleventh Edition has been completely updated and refreshed with an emphasis on state-of-the-art, evidence-based surgical care. You will find an exciting array of new contributors from around the world, new chapters on cutting-edge topics, plus the acclaimed learning aids that make the material easier to understand and memorize. This outstanding content is bolstered by more than 800 photographs and 1,300 line drawings, most in full color, as well as online videos demonstrating key operations. Here’s why the Eleventh Edition is the best edition yet: Six timely new chapters on important topics such as enhanced recovery after surgery (ERAS), ambulatory/outpatient surgery, evidence for surgery practice, skills and simulation, and web-based education and social media High-quality full-color design showcases an unsurpassed illustration program Emphasis on high-yield discussion of diagnosis and treatment of surgical disease, arranged by organ system and surgical specialty Acclaimed learning aids (many new to this edition), including an abundance of completely up-to-date tables that summarize the most current evidence, boxed key points, detailed anatomical figures, diagnostic and management algorithms, and an abundance of completely up-to-date tables, and key references More than the field’s cornerstone textbook, Schwartz’s Principles of Surgery is an international compendium of the knowledge and technique of the world’s leading surgeons.

CRISPR Cas9 mediated Somatic Correction of a Novel Coagulator Factor IX Gene Mutation Ameliorates Hemophilia in Mouse

CRISPR Cas9   mediated Somatic Correction of a Novel Coagulator Factor IX Gene Mutation Ameliorates Hemophilia in Mouse Book
Author : N.A
Publisher :
Release : 2016
ISBN :
Language : En, Es, Fr & De

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Book Description :

Abstract: The X‐linked genetic bleeding disorder caused by deficiency of coagulator factor IX, hemophilia B, is a disease ideally suited for gene therapy with genome editing technology. Here, we identify a family with hemophilia B carrying a novel mutation, Y371D, in the human F9 gene. The CRISPR/Cas9 system was used to generate distinct genetically modified mouse models and confirmed that the novel Y371D mutation resulted in a more severe hemophilia B phenotype than the previously identified Y371S mutation. To develop therapeutic strategies targeting this mutation, we subsequently compared naked DNA constructs versus adenoviral vectors to deliver Cas9 components targeting the F9 Y371D mutation in adult mice. After treatment, hemophilia B mice receiving naked DNA constructs exhibited correction of over 0.56% of F9 alleles in hepatocytes, which was sufficient to restore hemostasis. In contrast, the adenoviral delivery system resulted in a higher corrective efficiency but no therapeutic effects due to severe hepatic toxicity. Our studies suggest that CRISPR/Cas‐mediated in situ genome editing could be a feasible therapeutic strategy for human hereditary diseases, although an efficient and clinically relevant delivery system is required for further clinical studies. Synopsis: CRISPR/Cas9‐mediated genome editing holds promise for the treatment of genetic disorders, but its potential for hemophilia treatment is unknown. This study shows that in genome correction via Cas9 is a feasible therapeutic strategy for hemophilia B. Identification a family with hemophilia B carrying a novel mutation, Y371D, in the human F9 gene. Generation of three distinct genetically modified mouse models and confirmation that the mouse harboring the novel Y371D mutation is a new hemophilia B model. Hepatic in situ correction of the point mutation in the F9 allele via CRISPR/Cas9‐mediated genome editing was sufficient to restore hemostasis in hemophilia B mice. Abstract : CRISPR/Cas9‐mediated genome editing holds promise for the treatment of genetic disorders, but its potential for hemophilia treatment is unknown. This study shows that in genome correction via Cas9 is a feasible therapeutic strategy for hemophilia B.

Engineering Agrobacterium Tumefaciens to Facilitate and Improve Knock in Efficiency in Plants Via Cas9 Editing Strategies

Engineering Agrobacterium Tumefaciens to Facilitate and Improve Knock in Efficiency in Plants Via Cas9 Editing Strategies Book
Author : Adam John Kirby
Publisher :
Release : 2020
ISBN :
Language : En, Es, Fr & De

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Book Description :

The development of efficient tools for plant genome engineering is key to dramatically expedite basic research approaches and, at the same time, facilitate translational and biotechnological strategies in agriculture to improve crops and meet future food demand. For decades, technologies enabling precise Gene Targeting (GT) and efficient DNA knock-in or sequence replacement via Homology-Directed Repair (HDR) in plants has remained challenging, and although some recent reports suggest that progress is being made, no efficient and reproducible protocols have yet been established. Agrobacterium tumefaciens-mediated insertion entails the mobilization of the T-DNA into the host plant genome. However, this process occurs randomly and the system is very inefficient for targeted knock-ins. For this project, we capitalize on the extensive knowledge of Agrobacterium tumefaciens biology and combine it with CRISPR-Cas9 genome editing technologies to develop a novel strategy for high efficiency targeted knock-in. To do so, we decided to engineer Agrobacterium tumefaciens to help us reduce the distance between the T-DNA (harboring the cassette for knock-in) and the Cas9 cut site that marks the location where specifically the targeted insertion will occur. The set of tools we are presenting here will serve not to only accelerate basic research but also assist in engineering crop genomes in order to meet our future food demand in an efficient and sustainable manner.

Improving Fungal Genome Editing and Plant Disease Diagnostics with CRISPR Cas Technologies

Improving Fungal Genome Editing and Plant Disease Diagnostics with CRISPR Cas Technologies Book
Author : Matthew Wheatley
Publisher :
Release : 2020
ISBN :
Language : En, Es, Fr & De

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Book Description :

As a plant pathologist, I am interested in exploring new technologies to study plant-microbe interactions as well as improving crop disease management. Recently, the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein) system has emerged as a powerful technology with versatile applications in basic and applied research in medicine and agriculture. The goal of my dissertation project has been focused on improving and applying the CRISPR/Cas toolkit to facilitate efficient genome editing of fungal plant pathogens as well as sensitive and reliable diagnosis of plant diseases. The success of CRISPR/Cas9-based multiplex genome editing in the rice blast fungus and development of Cas12a-based diagnostics for citrus greening pathogen and phytoplasmas highlighted in my study demonstrate the utility and broad application of CRISPR/Cas technologies in plant pathology and disease management. The first objective of my research was to generate an online tool to aid in the design and selection of specific guide RNA (gRNA) spacers for genome editing of plant pathogens. The use of highly specific gRNAs is required to prevent unintended off-targeting effects. The CRISPR-PLANT v2 gRNA prediction pipeline was used for the genome-wide prediction of highly specific gRNA spacers for fifteen genomes of bacterial, oomycete, and fungal plant pathogens. Of the gRNAs predicted across these genomes, over 90% of the gRNA spacers belong to the highly specific classes of gRNA spacers and exhibit genome-wide targetability. The resulting gRNA spacer database and CRISPR-Pathogen webtool will be available to the plant pathology and microbiology research communities to facilitate the design of specific gRNAs and application of CRISPR/Cas9 genome editing in microbial plant pathogens. My second objective was to utilize the CRISPR/Cas9 system to improve the efficiency of single and multiplex genome editing in fungal plant pathogens. Magnaporthe oryzae is the causal agent of rice and wheat blast diseases and poses a major threat to rice and wheat production worldwide. As described in Chapter 3, CRISPR/Cas9-mediated multiplex genome editing in M. oryzae was successfully achieved using the polycistronic tRNA-gRNA (PTG) strategy. Upon creation of double stranded breaks (DSBs) by Cas9, targeted gene mutation in M. oryzae was created via either non-homologous end joining (NHEJ) or homology-directed repair (HDR) depending on the targeted loci. In absence of donor templates, Cas9-induced DSBs frequently triggered genomic rearrangement, leading to the loss of PCR-based amplification of target sites. By providing donor templates, however, HDR-mediated precise genome editing was achieved at the efficiency up to 100% when targeting a single locus. The PTG-based multiplex genome editing via HDR also successfully generated double and triple gene mutants. Interestingly, the HDR and NHEJ editing frequencies in M. oryzae appear to be dependent on genomic location of target sites and are likely influenced by flanking repetitive sequences and transposable elements. The resulting CRISPR/Cas9 tools and strategies from this study are expected to aid in the efficient genome editing and functional genomics analysis for M. oryzae and other fungal species. My third objective was to adapt the Cas12a-based method to enable highly sensitive and specific detection of pathogen nucleic acids for rapid and accurate diagnosis. As illustrated in Chapters 4 and 5, the citrus greening pathogen (Candidatus Liberibacter asiaticus) and purple potato top (PPT) phytoplasma were selected as target pathogens. The Cas12a-based DETECTR (DNA endonuclease-targeted CRISPR trans reporter) assay enabled highly specific and sensitive detection of CLas and Group VI phytoplasmas known to cause PPT from the infected samples. The DETECTR assay couples isothermal amplification and Cas12a trans-cleavage of fluorescent reporter oligos and enables detection of pathogen nucleic acids at the attomolar level. The DETECTR assay was able to accurately detect the presence of pathogen nucleic acids across all infected samples and was shown to be highly specific across closely related species. The improvement in detection sensitivity and flexibility of the DETECTR technology, positions the DETECTR assay as a suitable tool for early detection of pathogen nucleic acids. Furthermore, the DETECTR strategy allows flexibility to capture assay outputs with fluorescent microplate reader or lateral flow assay for potentially high-throughput and/or field-deployable disease diagnostics.

Genome Editing in Kenaf

Genome Editing in Kenaf Book
Author : Kangqi Li
Publisher :
Release : 2020
ISBN :
Language : En, Es, Fr & De

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Book Description :

The potential for Hibiscus cannabinus L. (kenaf) improvement via genome editing using the CRISPR/Cas9 system to generate gene knock-outs was explored. Studies included target gene identification, target guide RNA (gRNA) selection, plant tissue (explant) choice and media composition for plant regeneration. A putative kenaf phytoene desaturase gene (pds, GEED01047592.1) was identified in the kenaf transcriptome, and molecularly confirmed. Kenaf seedling tissues were transformed via Agrobacterium tumefaciens containing the cas9 gene (endonuclease required for gene knock-out) and each gRNA separately; putative transgenic calli and adventitious shoots arose on a medium containing 1-naphthaleneacetic acid, thidiazuron and silver nitrate. Tissues appeared chlorotic/albino and shoots remained diminutive/dwarf-like. These unique morphologies had also been noted by researchers who successfully knocked out the pds gene in other plant species. Cas9 DNA was detected in these putative transgenic kenaf tissues, but initial DNA sequencing analysis did not confirm knock-out/mutations in targeted areas of the pds gene.

Goodman and Gilman s The Pharmacological Basis of Therapeutics 13th Edition

Goodman and Gilman s The Pharmacological Basis of Therapeutics  13th Edition Book
Author : Laurence Brunton,Bjorn Knollman,Randa Hilal-Dandan
Publisher : McGraw Hill Professional
Release : 2017-10-26
ISBN : 1259584747
Language : En, Es, Fr & De

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Book Description :

The gold-standard of pharmacology texts – completely updated to reflect the latest research and developments A Doody’s Core Title for 2019! Goodman & Gilman’s: The Pharmacological Basis of Therapeutics, Thirteenth Edition represents the pinnacle of authority and accuracy in describing the actions and uses of therapeutic agents in relation to physiology and pathophysiology. Goodman & Gilman’s careful balance of basic science and clinical application has guided thousands of practitioners and students to a clear understanding of the drugs essential to preventing, diagnosing, and treating disease. The Thirteenth Edition includes more than 500 color illustrations, with many new figures emphasizing mechanisms of drug action. More than 30 new contributors have added to this edition, while the focus on basic principles is undiminished. This edition is enhanced by timely new content: •NEW chapters including Treatment of Pulmonary Arterial Hypertension, Immunity and Inflammation, Immunoglobulins and Vaccines, and Treatment of Viral Hepatitis •Expanded coverage of cardiovascular disease, with separate chapters on myocardial ischemia, hypertension, and heart failure •Increased emphasis on cellular signaling pathways involved in drug action •Summary tables at the end of each chapter that organize drugs discussed in that chapter into relevant categories and detail therapeutic usage, clinical pharmacology, and tips •Chapter Content Outlines at the beginning of each chapter •Abbreviation boxes in every chapter to easily identify the abbreviations appearing in that chapter More than a textbook, Goodman & Gilman’s is a working template for the effective and rational prescribing of drugs in daily practice.

Cas9 Ribonucleoprotein Delivery Via Microfluidic Cell Deformation Chip for Human T Cell Genome Editing and Immunotherapy

Cas9 Ribonucleoprotein Delivery Via Microfluidic Cell   Deformation Chip for Human T   Cell Genome Editing and Immunotherapy Book
Author : N.A
Publisher :
Release : 2017
ISBN :
Language : En, Es, Fr & De

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Book Description :

Abstract : This study reports a microfluidic cell deformation‐based method to deliver the Cas9 ribonucleoprotein (RNP) complexes to different cell types for efficient genome editing, including hard‐to‐transfect human primary CD4+ T cells. The RNP based CRISPR‐Cas9 system has great advantage in shortening reaction time and reducing off‐target problems, which holds great potential in future gene therapy applications.

Cell Culture Engineering

Cell Culture Engineering Book
Author : Gyun Min Lee,Helene Faustrup Kildegaard
Publisher : John Wiley & Sons
Release : 2020-01-13
ISBN : 3527343342
Language : En, Es, Fr & De

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Book Description :

Offers a comprehensive overview of cell culture engineering, providing insight into cell engineering, systems biology approaches and processing technology In Cell Culture Engineering: Recombinant Protein Production, editors Gyun Min Lee and Helene Faustrup Kildegaard assemble top class authors to present expert coverage of topics such as: cell line development for therapeutic protein production; development of a transient gene expression upstream platform; and CHO synthetic biology. They provide readers with everything they need to know about enhancing product and bioprocess attributes using genome-scale models of CHO metabolism; omics data and mammalian systems biotechnology; perfusion culture; and much more. This all-new, up-to-date reference covers all of the important aspects of cell culture engineering, including cell engineering, system biology approaches, and processing technology. It describes the challenges in cell line development and cell engineering, e.g. via gene editing tools like CRISPR/Cas9 and with the aim to engineer glycosylation patterns. Furthermore, it gives an overview about synthetic biology approaches applied to cell culture engineering and elaborates the use of CHO cells as common cell line for protein production. In addition, the book discusses the most important aspects of production processes, including cell culture media, batch, fed-batch, and perfusion processes as well as process analytical technology, quality by design, and scale down models. -Covers key elements of cell culture engineering applied to the production of recombinant proteins for therapeutic use -Focuses on mammalian and animal cells to help highlight synthetic and systems biology approaches to cell culture engineering, exemplified by the widely used CHO cell line -Part of the renowned "Advanced Biotechnology" book series Cell Culture Engineering: Recombinant Protein Production will appeal to biotechnologists, bioengineers, life scientists, chemical engineers, and PhD students in the life sciences.

Epigenetic Shaping of Sociosexual Interactions From Plants to Humans

Epigenetic Shaping of Sociosexual Interactions  From Plants to Humans Book
Author : N.A
Publisher : Academic Press
Release : 2014-09-03
ISBN : 9780128002223
Language : En, Es, Fr & De

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Book Description :

Epigenetic Shaping of Sociosexual Interactions: From Plants to Humans is the first attempt to interpret the higher social functions of organisms. This volume covers an extraordinarily wide range of biological research and provides a novel framework for understanding human-specific brain functions. Covers an extraordinarily wide range of biological research Provides a novel framework for understanding human-specific brain functions.

Development and Application of Advanced Synthetic Biology Tools for Engineering Chemical Production in Yarrowia Lipolytica

Development and Application of Advanced Synthetic Biology Tools for Engineering Chemical Production in Yarrowia Lipolytica Book
Author : Cory Schwartz
Publisher :
Release : 2018
ISBN : 9780438896932
Language : En, Es, Fr & De

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Book Description :

Renewable chemical production via microbial fermentation is a growing and critical industry. To overcome limitations in the productivity of commonly used hosts, nonconventional microbes that have uniquely advantageous metabolisms and phenotypes are needed. One such organism is the yeast Yarrowia lipolytica, which has a high native capacity to synthesize lipids and grow on a range of substrates. A significant bottleneck in engineering Y. lipolytica is a lack of synthetic biology tools for multiplexed genome editing and rapid strain development. To overcome these limitations, we have developed CRISPR-Cas9 based tools for (i) targeted gene disruption, (ii) expression cassette integration into predefined genomic loci, (iii) repression of native genes using CRISPR interference, (iv) activation of cryptic genes using CRISPR activation, and (v) genome-wide knockout screening. Straightforward adaptation of CRISPR-Cas9 strategies used in other eukaryotes had limited success, resulting in low gene disruption rates in Y. lipolytica. To improve this, we designed novel synthetic RNA polymerase III promoters for sgRNA expression, the best of which achieved gene disruption rates>90% and genome integration rates>50%. CRISPR-mediated markerless genome integration was used to make a strain of Y. lipolytica capable of producing over 21 mg/g DCW of the valuable carotenoid lycopene. We also adapted the CRISPR-Cas9 system for gene repression and significantly increased homologous recombination by transiently repressing genes involved in nonhomologous end-joining. The system was further adapted for gene activation and used to express natively silent [Beta]-glucosidase genes to enable growth on cellobiose. To accelerate strain engineering, we designed and constructed a library of plasmids expressing ~48,000 sgRNAs that target all protein coding sequences in the genome with 6-fold coverage. By transforming this pooled library into Y. lipolytica strains expressing Cas9 and quantifying sgRNA enrichment or depletion after outgrowth, genes important for growth under different conditions can be identified. By performing screening experiments in different strain backgrounds, determinants of CRISPR-Cas9 function can be characterized and industrially relevant phenotypes can be selected for.

Efficient and Multiplex Genome Editing Using GeneArt CRISPR Nuclease MRNA

Efficient and Multiplex Genome Editing Using GeneArt   CRISPR Nuclease MRNA Book
Author : Johan Garza,Jon Chesnut,Sanjay Kumar,Ravinder Namritha,Shantanu Kumar
Publisher :
Release : 2015
ISBN :
Language : En, Es, Fr & De

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Book Description :

CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system is derived from a prokaryotic adaptive immune system that uses an RNA-guided DNA nuclease to mutate and silence viral DNA infections (Jinek et al., 2012). These nucleases have been re-engineered to function as gene editing tools in various organisms, including mammalian cells (Cong et al., 2013). Thermo Fisher Scientific has developed the GeneArt® CRISPR Nuclease mRNA and in vitro transcribed gRNA from GeneArt® T7 string DNA that function together to efficiently and conveniently modify the genomic loci of choice, while circumventing promoter-associated restrictions, avoiding random DNA integrations from previous DNA vector-based solutions and show potential for in vivo applications. The goal of the Biotech Master's Semester In Residence (SIR) project is to demonstrate product and application data of GeneArt® CRISPR Nuclease mRNA (Cas9 mRNA) system coupled with in vitro transcribed (IVT) guide RNAs (gRNAs) showing efficient editing in a broad range of cell types including human and mouse derived cell lines, as well as induced pluripotent stem cells (iPSCs). Further, to demonstrate its capabilities, we used GeneArt® CRISPR nuclease mRNA and a set of gRNAs to simultaneously target several genomic loci at once in a multiplexed fashion and to result in clonal populations with mutations at chosen loci. Such clones were screened using the gel-based Genomic Cleavage Detection assay and confirmed through Sanger sequencing of PCR amplified targeted DNA fragments via targeted DNA via TOPO® TA Cloning kit. The report within demonstrates how the GeneArt® CRISPR nuclease mRNA (Catalog # A25640) can be used for performing CRISPR-Cas9 mediated genome editing. Coupled with IVT gRNA, where one can efficiently and conveniently modify their target genomic sequences. Because of eliminating promoter-associated constraints, the complete RNA format combined with RNA-specific transfection reagents, provides superior delivery combined with high genome editing efficiency in broad cell types including stem cells. GeneArt® CRISPR Nuclease mRNA system along with the complete work flow solutions described in the report can address both small and large scale cell engineering needs.

SaCas9 Requires 5 NNGRRT 3 PAM for Sufficient Cleavage and Possesses Higher Cleavage Activity Than SpCas9 Or FnCpf1 in Human Cells

SaCas9 Requires 5      NNGRRT   3    PAM for Sufficient Cleavage and Possesses Higher Cleavage Activity Than SpCas9 Or FnCpf1 in Human Cells Book
Author : N.A
Publisher :
Release : 2018
ISBN :
Language : En, Es, Fr & De

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Book Description :

Abstract: CRISPR/Cas9‐mediated gene therapy holds great promise for the treatment of human diseases. The protospacer adjacent motif (PAM), the sequence adjacent to the target sequence, is an essential targeting component for the design of CRISPR/Cas9‐mediated gene editing. However, currently, very few studies have attempted to directly study the PAM sequence in human cells. To address this issue, the authors develop a dual fluorescence reporter system that could be harnessed for identifying functional PAMs for genome editing endonuclease, including Cas9. With this system, the authors investigate the effects of different PAM sequences for SaCas9, which is small and has the advantage of allowing in vivo genome editing, and found only 5′‐NNGRRT‐3′ PAM could induced sufficient target cleavage with multi‐sites. The authors also found SaCas9 possesses higher activity than SpCas9 or FnCpf1 via plasmids (episomal) and chromosomes with integrated eGFP‐based comparison. Taken together, the authors show that a dual fluorescence reporter system is a means to identifying a functional PAM and quantitatively comparing the efficiency of different genome editing endonucleases with the similar or identical target sequence in human cells. Abstract : Currently, very few studies have attempted to study the PAM sequence of CRISPR/Cas9 directly in human cells. To address this issue, the authors develop a dual fluorescence reporter system (pmT/mG), and found SaCas9 requires 5′‐NNGRRT‐3' for sufficient cleavage in human cells. The authors also found SaCas9 possesses higher activity than SpCas9 or FnCpf1 in human cells via plasmids (episomal) and chromosomes based comparison.

Harpers Illustrated Biochemistry 30th Edition

Harpers Illustrated Biochemistry 30th Edition Book
Author : Victor W. Rodwell,David Bender,Kathleen M. Botham,Peter J. Kennelly,P. Anthony Weil
Publisher : McGraw Hill Professional
Release : 2015-03-22
ISBN : 0071825371
Language : En, Es, Fr & De

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Book Description :

Gain a thorough understanding of the principles ofbiochemistry as they relate to the study of clinical medicine A Doody's Core Title for 2017! THE BEST REVIEW FOR THE USMLE! The Thirtieth Edition of Harper’s Illustrated Biochemistry combines outstanding full-colorillustrations with authoritative integrated coverage of biochemical disease and clinical information. Using brevity and numerous medically relevant examples, Harper's presents a clear, succinct review of the fundamentals of biochemistry that every student must understand in order to succeed in medical school. All fifty-eight chapters emphasize the medical relevance of biochemistry Full-color presentation includes more than 600 illustrations Each chapter includes a section on BiomedicalImportance and a summary of the topics covered Review questions follow each of the eleven sections Case studies in every chapter emphasize the clinical relevance to biochemistry NEW coverage of toxic naturally-occurring amino acids; extraterrestrial biomolecules; computer-aided drug design; the role of complement cascade in bacterial and viral infection; secreted mediators of cell-cell signaling between leukocytes; the role of mast cells, basophils, andeosinophils; and the hazard of antioxidants that down-regulate radical signaling for apoptosis and increase risk of cancer Applauded by medical students for its current and engaging style, Harper's Illustrated Biochemistry is an essential for USMLE review and the single best reference for learning the clinical relevance of any biochemistry topic.

Williams Hematology 9E

Williams Hematology  9E Book
Author : Kenneth Kaushansky,Marshall A. Lichtman,Josef Prchal,Marcel M. Levi,Oliver W Press,Linda J Burns,Michael Caligiuri
Publisher : McGraw Hill Professional
Release : 2015-12-23
ISBN : 0071833013
Language : En, Es, Fr & De

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Book Description :

Publisher's Note: Products purchased from Third Party sellers are not guaranteed by the publisher for quality, authenticity, or access to any online entitlements included with the product. The world's most highly regarded reference text on the mechanisms and clinical management of blood diseases A Doody's Core Title for 2019! Edition after edition, Williams Hematology has guided generations of clinicians, biomedical researchers, and trainees in many disciplines through the origins, pathophysiological mechanisms, and management of benign and malignant disorders of blood cells and coagulation proteins. It is acknowledged worldwide as the leading hematology resource, with editors who are internationally regarded for their research and clinical achievements and authors who are luminaries in their fields. The Ninth Edition of Williams Hematology is extensively revised to reflect the latest advancements in basic science, translational pathophysiology, and clinical practice. In addition to completely new chapters, it features a full-color presentation that includes 700 photographs, 300 of which are new to this edition, and 475 illustrations. Recognizing that blood and marrow cell morphology is at the heart of diagnostic hematology, informative color images of the relevant disease topics are conveniently integrated into each chapter, allowing easy access to illustrations of cell morphology important to diagnosis. Comprehensive in its depth and breath, this go-to textbook begins with the evaluation of the patient and progresses to the molecular and cellular underpinnings of normal and pathological hematology. Subsequent sections present disorders of the erythrocyte, granulocytes and monocytes, lymphocytes and plasma cells, malignant myeloid and lymphoid diseases, hemostasis and thrombosis, and transfusion medicine.

Biology of Brain Disorders

Biology of Brain Disorders Book
Author : Daniela Tropea,Andrew Harkin
Publisher : Frontiers Media SA
Release : 2018-01-31
ISBN : 2889453804
Language : En, Es, Fr & De

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Book Description :

Brain disorders, including neurological and neuropsychiatric conditions, represent a challenge for public health systems and society at large. The limited knowledge of their biology hampers the development of diagnostic tools and effective therapeutics. A clear understanding of the mechanisms that underlie the onset and progression of brain disorders is required in order to identify new avenues for therapeutic intervention. Overlapping genetic risk factors across different brain disorders suggest common linkages and pathophysiological mechanisms that underlie brain disorders. Methodological and technological advances are leading to new insights that go beyond traditional hypotheses. Taking account of underlying molecular, cellular and systems biology underlying brain function will play an important role in the classification of brain disorders in future. In this Research Topic, the latest advances in our understanding of biological mechanisms across different brain disorders are presented. The areas covered include developments in neurogenetics, epigenetics, plasticity, glial cell biology, neuroimmune interactions and new technologies associated with the study of brain function. Examples of how understanding of biological mechanisms are translating into research strategies that aim to advance diagnoses and treatment of brain disorders are discussed.