Download or read book Sophisticated Mammalian Cell Engineering with CRISPR Cas12a written by Hannah Rose Kempton and published by . This book was released on 2022 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: The application of CRISPR/Cas systems for programmable DNA manipulation has revolutionized the field of genome engineering. In addition to their use for cutting DNA, nuclease-dead versions of Cas proteins fused to different effectors are a powerful tool for cell engineering and synthetic biology. In this thesis, I will discuss two projects from my Ph.D., both demonstrating how Cas12a can be used for constructing next-generation genetic circuits in human cells. For my first project I developed a split Cas12a protein, which spontaneously re-assembles into a functional gene-activating effector when all components are present. Therapeutic cell engineering, in which cells are programmed to sense and respond to complex disease states, requires integration of information about the cellular environment and execution of a functional response only in response to the specified combination of inputs. By linking the expression of individual components of the split Cas12a system to different input signals, it forms the functional core of multi-input, multi-output logic circuits in mammalian cells. The system is highly programmable and can generate expandable AND gates, as well as incorporate NOT logic by using anti-CRISPR proteins as an OFF switch. For my second project I established a platform for parallel signal recording using a hyper-efficient Cas12a base editor. Biological signal recording enables the study of molecular inputs experienced throughout cellular history, by encoding a transient signal as a permanent, targeted DNA mutation that can be read out at a later time by sequencing. However, current methods are often limited in their ability to scale up beyond a single biologically relevant signal in mammalian contexts. This novel implementation of Cas12a for signal recording provides a powerful platform for scalable recording, demonstrated by the recording of up to four signals in parallel in human cells. This method is plug-and-play with diverse inputs in different cell types, including tumor antigen recording in CAR-T cells. It can also be easily extended for more sophisticated applications, such as time-delimited recording and dual recorder-effector cells. Overall, these two projects demonstrate the utility of Cas-based effectors in complex genetic circuits, for both understand-ing fundamental biological processes as well as constructing smarter cellular therapeutics.
Download or read book CRISPR Cas Systems written by Rodolphe Barrangou and published by Springer Science & Business Media. This book was released on 2012-12-13 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt: CRISPR/Cas is a recently described defense system that protects bacteria and archaea against invasion by mobile genetic elements such as viruses and plasmids. A wide spectrum of distinct CRISPR/Cas systems has been identified in at least half of the available prokaryotic genomes. On-going structural and functional analyses have resulted in a far greater insight into the functions and possible applications of these systems, although many secrets remain to be discovered. In this book, experts summarize the state of the art in this exciting field.
Download or read book Mammalian Cell Engineering written by Ryosuke Kojima and published by Humana. This book was released on 2022-07-21 with total page 330 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume explores the latest engineering methods of mammalian cells that are useful for controlling the performance of engineered mammalian cells for future cell-based therapeutics and for better understanding of complex biological systems. The chapters in this book are organized into five parts. Part One described methods to engineer mammalian cells to sense biologically relevant inputs, such as cell contacts and soluble proteins. Part Two looks at techniques to engineer mammalian cells to sense artificial inputs, such as light and ultrasound. Part Three provides cutting-edge CRISPR-Cas-based methods to carry out highly multiplexed genome editing and spatiotemporally controlled genome editing. Part Four discusses ways to control and engineer biological events in mammalian cells in combination with chemical compounds and systems. Part Five explores techniques to engineer specific mammalian cells in targeted manners. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and authoritative, Mammalian Cell Engineering: Methods and Protocols is a valuable resource that allows scientists to successfully carry out their research, thus ultimately contributing to the future advancement of this field.
Download or read book CRISPR Cas12a assisted Gene Tagging in Mammalian Cells written by Krisztina Gubicza and published by . This book was released on 2023 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book CRISPR Cas Enzymes written by and published by Academic Press. This book was released on 2019-01-25 with total page 454 pages. Available in PDF, EPUB and Kindle. Book excerpt: CRISPR-Cas Enzymes, Volume 616, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Topics covered in this release include CRISPR bioinformatics, A method for one-step assembly of Class 2 CRISPR arrays, Biochemical reconstitution and structural analysis of ribonucleoprotein complexes in Type I-E CRISPR-Cas systems, Mechanistic dissection of the CRISPR interference pathway in Type I-E CRISPR-Cas system, Site-specific fluorescent labeling of individual proteins within CRISPR complexes, Fluorescence-based methods for measuring target interference by CRISPR-Cas systems, Native State Structural Characterization of CRISRP Associated Complexes using Mass Spectrometry, and more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Enzymology series - Updated release includes the latest information on the CRISPR-Cas Enzymes
Download or read book RNA Nanotechnology written by Bin Wang and published by CRC Press. This book was released on 2014-04-02 with total page 468 pages. Available in PDF, EPUB and Kindle. Book excerpt: In the past few decades there has been incredible growth in "bionano"-related research, which has been accompanied by numerous publications in this field. Although various compilations address topics related to deoxyribonucleic acid (DNA) and protein, there are few books that focus on determining the structure of ribonucleic acid (RNA) and using RNA as building blocks to construct nanoarchitectures for biomedical and healthcare applications. RNA Nanotechnology is a comprehensive volume that details both the traditional approaches and the latest developments in the field of RNA-related technology. This book targets a wide audience: a broad introduction provides a solid academic background for students, researchers, and scientists who are unfamiliar with the subject, while the in-depth descriptions and discussions are useful for advanced professionals. The book opens with reviews on the basic aspects of RNA biology, computational approaches for predicting RNA structures, and traditional and emerging experimental approaches for probing RNA structures. This section is followed by explorations of the latest research and discoveries in RNA nanotechnology, including the design and construction of RNA-based nanostructures. The final segment of the book includes descriptions and discussions of the potential biological and therapeutic applications of small RNA molecules, such as small/short interfering RNAs (siRNAs), microRNAs (miRNAs), RNA aptamers, and ribozymes.
Download or read book Mammalian Synthetic Biology written by Jamie Davies and published by . This book was released on 2019-12-12 with total page 137 pages. Available in PDF, EPUB and Kindle. Book excerpt: Written primarily for mid-to-upper level undergraduates, this primer will introduce students to topics at the forefront of the subject that are being applied to probe biological problems, or to address the most pressing issues facing society. These topics will include those that form thecornerstone of contemporary research, helping students to make the transition to active researcher.This primer introduces the challenges and opportunities of applying synthetic biological techniques to mammalian cells, tissues, and organisms. It covers the special features that make engineering mammalian systems different from engineering bacteria, fungi, and plants, and provides an overview ofcurrent techniques. A variety of cutting-edge examples illustrate the different purposes of mammalian synthetic biology, including pure biomedical research, drug production, tissue engineering, and regenerative medicine.
Download or read book Capillary Gel Electrophoresis written by Andras Guttman and published by Newnes. This book was released on 2021-12-04 with total page 391 pages. Available in PDF, EPUB and Kindle. Book excerpt: Capillary Gel Electrophoresis and Related Microseparation Techniques covers all theoretical and practical aspects of capillary gel electrophoresis. It also provides an excellent overview of the key application areas of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods. It not only gives readers a better understanding of how to utilize this technology, but also provides insights into how to determine which method will provide the best technical solutions to particular problems. This book can also serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses. - Covers all theoretical and practical aspects of capillary gel electrophoresis - Excellent overview of the key applications of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods - Teaches readers how to use the technology and select methods that are ideal for fundamental problems - Can serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses
Download or read book Gene Editing written by Yuan-Chuan Chen and published by BoD – Books on Demand. This book was released on 2019-05-29 with total page 110 pages. Available in PDF, EPUB and Kindle. Book excerpt: Gene-editing technologies (e.g., ZFNs, TALENs, and CRISPRs/Cas9) have been extensively used as tools in basic research. They are further applied in manufacturing agricultural products, food, industrial products, medicinal products, etc. Particularly, the discovery of medicinal products using gene-editing technologies will open a new era for human therapeutics. Though there are still many technical and ethical challenges ahead of us, more and more products based on gene-editing technologies have been approved for marketing. These technologies are promising for multiple applications. Their development and implications should be explored in the broadest context possible. Future research directions should also be highlighted. In this book, the applications, perspectives, and challenges of gene-editing technologies are significantly demonstrated and discussed.
Download or read book The CRISPR Cas System written by Muhammad Jamal and published by Caister Academic Press Limited. This book was released on 2017 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The use of CRISPR/Cas technology for genome editing suggests many potential applications, including the alteration of the germline of humans, animals and food crops. The speed and efficiency of the CRISPR/Cas system make it a potentially useful system for gene therapy. In this volume expert international authors provide a useful and timely review of the applications of the CRISPR/Cas system across diverse fields and explore further avenues and research directions of this novel and powerful editing technology. The technology and its application are reviewed with respect to reproduction and development, immunity and genetic diseases, system structure and system specificity. Some of the potential problems of the CRISPR/Cas system are also discussed, in particular the specificity of the system: this remains an important topic as improvement could lead to the more direct and efficient use of the CRISPR/Cas system in clinical settings. The authors also debate ethical concerns associated with this powerful new technology. This volume is a rigorous review of the applications and new opportunities for the CRISPR/Cas system and provides a stimulus for current and future research. An invaluable guide for all scientists working in the fields of genome editing and gene therapy the book is also recommended for all life sciences libraries.
Download or read book Gene Editing in Plants written by and published by Academic Press. This book was released on 2017-07-14 with total page 266 pages. Available in PDF, EPUB and Kindle. Book excerpt: Gene Editing in Plants, Volume 149 aims to provide the reader with an up-to-date survey of cutting-edge research with gene editing tools and an overview of the implications of this research on the nutritional quality of fruits, vegetables and grains. New chapters in the updated volume include topics relating to Genome Engineering and Agriculture: Opportunities and Challenges, the Use of CRISPR/Cas9 for Crop Improvement in Maize and Soybean, the Use of Zinc-Finger Nucleases for Crop Improvement, Gene Editing in Polyploid Crops: Wheat, Camelina, Canola, Potato, Cotton, Peanut, Sugar Cane, and Citrus, and Gene Editing With TALEN and CRISPR/Cas in Rice. This ongoing serial contain contributions from leading scientists and researchers in the field of gene editing in plants who describe the results of their own research in this rapidly expanding area of science. - Shows the importance of revolutionary gene editing technology on plant biology research and its application to agricultural production - Provides insight into what may lie ahead in this rapidly expanding area of plant research and development - Contains contributions from major leaders in the field of plant gene editing
Download or read book Directed Evolution of Selective Enzymes written by Manfred T. Reetz and published by John Wiley & Sons. This book was released on 2016-12-19 with total page 320 pages. Available in PDF, EPUB and Kindle. Book excerpt: Authored by one of the world's leading organic chemists, this authoritative reference provides an overview of basic strategies in directed evolution and introduces common gene mutagenesis, screening and selection methods. Throughout the text, emphasis is placed on methodology development to maximize efficiency, reliability and speed of the experiments and to provide guidelines for efficient protein engineering. Professor Reetz highlights the application of directed evolution experiments to address limitations in the field of enzyme selectivity, substrate scope, activity and robustness. He critically reviews recent developments and case studies, takes a look at future applications in the field of organic synthesis, and concludes with lessons learned from previous experiments.
Download or read book Heritable Human Genome Editing written by The Royal Society and published by National Academies Press. This book was released on 2021-01-16 with total page 239 pages. Available in PDF, EPUB and Kindle. Book excerpt: Heritable human genome editing - making changes to the genetic material of eggs, sperm, or any cells that lead to their development, including the cells of early embryos, and establishing a pregnancy - raises not only scientific and medical considerations but also a host of ethical, moral, and societal issues. Human embryos whose genomes have been edited should not be used to create a pregnancy until it is established that precise genomic changes can be made reliably and without introducing undesired changes - criteria that have not yet been met, says Heritable Human Genome Editing. From an international commission of the U.S. National Academy of Medicine, U.S. National Academy of Sciences, and the U.K.'s Royal Society, the report considers potential benefits, harms, and uncertainties associated with genome editing technologies and defines a translational pathway from rigorous preclinical research to initial clinical uses, should a country decide to permit such uses. The report specifies stringent preclinical and clinical requirements for establishing safety and efficacy, and for undertaking long-term monitoring of outcomes. Extensive national and international dialogue is needed before any country decides whether to permit clinical use of this technology, according to the report, which identifies essential elements of national and international scientific governance and oversight.
Download or read book Synthetic Biology written by Christina Smolke and published by John Wiley & Sons. This book was released on 2018-02-28 with total page 532 pages. Available in PDF, EPUB and Kindle. Book excerpt: A review of the interdisciplinary field of synthetic biology, from genome design to spatial engineering. Written by an international panel of experts, Synthetic Biology draws from various areas of research in biology and engineering and explores the current applications to provide an authoritative overview of this burgeoning field. The text reviews the synthesis of DNA and genome engineering and offers a discussion of the parts and devices that control protein expression and activity. The authors include information on the devices that support spatial engineering, RNA switches and explore the early applications of synthetic biology in protein synthesis, generation of pathway libraries, and immunotherapy. Filled with the most recent research, compelling discussions, and unique perspectives, Synthetic Biology offers an important resource for understanding how this new branch of science can improve on applications for industry or biological research.
Download or read book CRISPR Cas Methods written by M. Tofazzal Islam and published by Humana. This book was released on 2020-06-30 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume details the fundamentals of the CRISPR-Cas system, and its protocols illustrate advances in CRISPR-Cas techniques for efficient genome editing. Introductory chapters provide a wide horizon of CRISPR/Cas-based methods and applications. Additional chapters guide readers through HDR-mediated editing, sgRNA design, the step-by-step procedure of multiplex adenine base editing experiments in rice, generating mutants for rice, wheat, Brachypodium, Barley, Flax, and Phytophthora, visual screening of mutants, gene deletion (knock-out), tagging (knock-in) in mammalian cells, the cloning-free (DNA-free) technique, cell-penetrating peptides, generating a genome-edited banana, and nuclear genome editing of Chlamydomonas employing CRISPR-Cpf1 combined with a single-stranded DNA (ssODN) repair template. Authoritative and cutting-edge, CRISPR-Cas Methods aims to assist researchers who are new to the field and are aiming to learn how best to adopt this technology for a particular organism.
Download or read book CRISPR written by Rodolphe Barrangou and published by John Wiley & Sons. This book was released on 2022-09-14 with total page 308 pages. Available in PDF, EPUB and Kindle. Book excerpt: “An excellent compendium of all things CRISPR from some of the leading minds in the field. With thorough coverage from every angle and beautifully detailed illustrations, this book is not to be missed!” Jennifer A. Doudna, Professor of Chemistry, Biochemistry & Molecular Biology, UC Berkeley; Founder, Innovative Genomics Institute; Nobel Laureate and coinventor of CRISPR technology “This journey through CRISPR biology and several of its breakthrough applications offers an exciting glimpse into one of the most beautiful and compelling fields in the life sciences.” David R. Liu, Director of the Merkin Institute at the Broad Institute of MIT and Harvard; Professor of Chemistry and Chemical Biology at Harvard University; coinventor of base editing and prime editing “A must read! The CRISPR topics, written by world-leading experts, span from the fascinating mechanistic underpinnings to the ingenious applications. One can read from start to finish or pick and choose themes. Either way, the book delivers utterly enjoyable learning!” Bonnie Bassler, Squibb Professor and Chair, Princeton University Department of Molecular Biology; Howard Hughes Medical Institute Investigator CRISPR-Cas systems have revolutionized the science of gene editing and their possible applications continue to expand, from basic research to potentially groundbreaking medical and commercial uses. Led by a distinguished team of editors, CRISPR: Biology and Applications explores the subject matter needed to delve into this fascinating area. Topics covered include: Classification and molecular mechanisms of CRISPR-Cas systems CRISPR-Cas evolution, regulation, expression, and function Uses for gene editing and modulation of gene expression CRISPR-based antimicrobials and phage resistance for medical and industrial purposes Written by internationally renowned authors, CRISPR: Biology and Applications serves as both an introductory guide for those new to the field and an authoritative reference for seasoned researchers whose work touches this evolving and headline- making science.
Download or read book Genome Editing and Engineering written by Krishnarao Appasani and published by Cambridge University Press. This book was released on 2018-08-23 with total page 535 pages. Available in PDF, EPUB and Kindle. Book excerpt: A complete guide to endonuclease-based genomic engineering, from basic science to application in disease biology and clinical treatment.
