Evolutionary Consequences of a CRISPR/Cas9-based Gene Drive in Saccharomyces Cerevisiae PDF Download
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Author: Sean Edward Gomez Guy
Publisher:
ISBN:
Category :
Languages : en
Pages : 48
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Book Description
Vector-borne diseases pose a significant health threat to millions. Genetic modification through CRISPR/Cas9-based gene drive technologies may be able to solve this problem by rapidly spreading disease resistance throughout vector populations. However, the long-term effects of active gene drive elements at evolutionary scales has yet to be extensively explored. To address this issue, we constructed S. cerevisiae strains that express both Cas9 and gRNA, Cas9 and no gRNA, or neither Cas9 nor gRNA. We founded replicate populations and propagated these over 1000 generations. From whole-population, whole-genome sequences, we identified changes in allele frequency in each population. By comparing the numbers of alleles to appear or fix in each population, we find evidence of higher mutation rates in strains expressing Cas9 nuclease. In parallel, we directly measure the mutation rate differences due to gene drive or Cas9 alone. Our work underscores the need to control CRISPR/Cas9 activity to limit unintended effects in natural populations.
Author: Sean Edward Gomez Guy
Publisher:
ISBN:
Category :
Languages : en
Pages : 48
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Book Description
Vector-borne diseases pose a significant health threat to millions. Genetic modification through CRISPR/Cas9-based gene drive technologies may be able to solve this problem by rapidly spreading disease resistance throughout vector populations. However, the long-term effects of active gene drive elements at evolutionary scales has yet to be extensively explored. To address this issue, we constructed S. cerevisiae strains that express both Cas9 and gRNA, Cas9 and no gRNA, or neither Cas9 nor gRNA. We founded replicate populations and propagated these over 1000 generations. From whole-population, whole-genome sequences, we identified changes in allele frequency in each population. By comparing the numbers of alleles to appear or fix in each population, we find evidence of higher mutation rates in strains expressing Cas9 nuclease. In parallel, we directly measure the mutation rate differences due to gene drive or Cas9 alone. Our work underscores the need to control CRISPR/Cas9 activity to limit unintended effects in natural populations.
Author: Yao Yan
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Book Description
The budding yeast Saccharomyces cerevisiae is a well-studied eukaryotic model organism for investigating the fundamental aspects of molecular and cellular biology. It also can be used to evaluate the technical feasibility of new technologies in eukaryotic cells. In our lab, we aimed to use Saccharomyces cerevisiae as a model to develop CRISPR/Cas9-based gene drive biotechnology. The CRISPR/Cas9 biotechnology is a powerful gene editing tool used to modify target genomic sequences in all forms of living creatures including animals, plants, fungi, and bacteria. One potential application of this molecular method is within a "gene drive" system. This unique arrangement of CRISPR within a genome may one day allow for global control of biological populations and be used to eliminate pests, parasites, and invasive species. However, there are many concerns regarding the utilization of this technology, including gene drive design, control, and development of resistance, etc. In our study, we developed an artificial gene drive system in budding yeast, which could edit multiple loci at the same time. We demonstrated that this triple gene drive system could successfully edit three DNA targets independently with only a single copy of S. pyogenes Cas9. We also found that the occurrence of NHEJ could be repressed by modifying DNA Ligase IV. However, successful gene drives still allowed for the occurrence of a small number of "resistant" clones. We investigated potential causes of this imperfect drive activity. Our work illustrated that imperfect activation of the inducible promoter driving expression of the Cas9 nuclease or issues with multiplexing to artificial sequences may have resulted in a small percentage of resistant/inactive clones. The CRISPR/Cas9 system can also be used to regulate gene transcription. This involves a mutated Cas9 variant that has lost its nuclease activity (dCas9). We developed a CRISPR/dCas9 system by tagging dCas9 with transcriptional regulators. Our experiments demonstrated that CRISPR/dCas9 could activate target gene transcription when tagged with the transcriptional activator VPR and repress gene transcription when tagged with the transcriptional repressor Mxi1.
Author: National Academies of Sciences, Engineering, and Medicine
Publisher: National Academies Press
ISBN: 0309437873
Category : Science
Languages : en
Pages : 231
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Book Description
Research on gene drive systems is rapidly advancing. Many proposed applications of gene drive research aim to solve environmental and public health challenges, including the reduction of poverty and the burden of vector-borne diseases, such as malaria and dengue, which disproportionately impact low and middle income countries. However, due to their intrinsic qualities of rapid spread and irreversibility, gene drive systems raise many questions with respect to their safety relative to public and environmental health. Because gene drive systems are designed to alter the environments we share in ways that will be hard to anticipate and impossible to completely roll back, questions about the ethics surrounding use of this research are complex and will require very careful exploration. Gene Drives on the Horizon outlines the state of knowledge relative to the science, ethics, public engagement, and risk assessment as they pertain to research directions of gene drive systems and governance of the research process. This report offers principles for responsible practices of gene drive research and related applications for use by investigators, their institutions, the research funders, and regulators.
Author: L. David Sibley
Publisher: John Wiley & Sons
ISBN: 1118308131
Category : Science
Languages : en
Pages : 912
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Book Description
A unique and timely review of the emergence of eukaryotic virulence in fungi, oomycetes, and protozoa, as they affect both animals and plants Evolution of Virulence in Eukaryotic Microbes addresses new developments in defining the molecular basis of virulence in eukaryotic pathogens. By examining how pathogenic determinants have evolved in concert with their hosts, often overcoming innate and adaptive immune mechanisms, the book takes a fresh look at the selective processes that have shaped their evolution. Introductory chapters ground the reader in principal evolutionary themes such as phylogenetics and genetic exchange, building a basis of knowledge for later chapters covering advances in genetic tools, how pathogens exchange genetic material in nature, and the common themes of evolutionary adaptation that lead to disease in different hosts. With the goal of linking the research findings of the many disparate scientific communities in the field, the book: Assembles for the first time a collection of chapters on the diversity of eukaryotic microorganisms and the influence of evolutionary forces on the origins and emergence of their virulent attributes Highlights examples from three important, divergent groups of eukaryotic microorganisms that cause disease in animals and plants: oomycetes, protozoan parasites, and fungi Covers how the development of genetic tools has fostered the identification and functional analyses of virulence determinants Addresses how pathogens exchange genetic material in nature via classical or modified meiotic processes, horizontal gene transfer, and sexual cycles including those that are cryptic or even unisexual Provides a broad framework for formulating future studies by illustrating themes common to different pathogenic microbes Evolution of Virulence in Eukaryotic Microbes is an ideal book for microbiologists, evolutionary biologists and medical professionals, as well as graduate students, postdoctoral fellows, and faculty members working on the evolution of pathogens.
Author: Miguel Fernández-Niño
Publisher: BoD – Books on Demand
ISBN: 1839696389
Category : Science
Languages : en
Pages : 106
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Book Description
The current advances in sequencing, data mining, DNA synthesis, cloning, in silico modeling, and genome editing have opened a new field of research known as Synthetic Genomics. The main goal of this emerging area is to engineer entire synthetic genomes from scratch using pre-designed building blocks obtained by chemical synthesis and rational design. This has opened the possibility to further improve our understanding of genome fundamentals by considering the effect of the whole biological system on biological function. Moreover, the construction of non-natural biological systems has allowed us to explore novel biological functions so far not discovered in nature. This book summarizes the current state of Synthetic Genomics, providing relevant examples in this emerging field.
Author: Rodolphe Barrangou
Publisher: Springer Science & Business Media
ISBN: 364234657X
Category : Science
Languages : en
Pages : 300
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Book Description
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.
Author: Christina Smolke
Publisher: John Wiley & Sons
ISBN: 3527688099
Category : Science
Languages : en
Pages : 532
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Book Description
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.
Author: Jennifer A. Doudna
Publisher:
ISBN: 9781621821304
Category : Genetics
Languages : en
Pages : 0
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Book Description
CRISPR/Cas-based techniques are revolutionizing the way geneticists and molecular biologists modify DNA sequences and modulate gene expression in cells and organisms. This laboratory manual presents step-by-step protocols for applying this cutting-edge technology to any system of interest. Contributors describe approaches for de.
Author: Zach N. Adelman
Publisher: Academic Press
ISBN: 0128004053
Category : Medical
Languages : en
Pages : 487
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Book Description
Genetic Control of Malaria and Dengue focuses on the knowledge, technology, regulation and ethics of using genetically modified mosquitoes to interrupt the transmission of important vector-borne diseases including Malaria. It contains coverage of the current state of knowledge of vector-borne diseases and how they are currently controlled; vaccine, drug and insecticide development; various strategies for altering the genome of mosquitoes in beneficial ways; and the regulatory, ethical and social environment concerning these strategies. For more than five decades, the prospect of using genetically-modified mosquitoes to control vector-borne disease transmission has been a purely hypothetical scenario. We simply did not have the technology or basic knowledge to be able to do it. With the explosion of field trials and potential interventions in development, Genetic Control of Malaria and Dengue provides a comprehensive overview of research in genetics, microbiology, virology, and ecology involved in the development and implementation of genetic modification programs for virus and disease control. This book is meant to provide a practical guide to researchers, regulators and the general public about how this technology actually works, how it can be improved, and what is still unknown. Includes coverage of vectorial capacity, critical to understanding vector-borne disease transmission Provides a summary of the concepts of both population suppression and population replacement Contains pivotal coverage of ethical and ecological ramifications of genetics-based control strategies
Author:
Publisher: Academic Press
ISBN: 0128125071
Category : Science
Languages : en
Pages : 150
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Book Description
CRISPR in Animals and Animal Models, Volume 152, the latest release in the Progress in Molecular Biology and Translational Science series, explores the genome editing CRISPR system in cells and animal models, its applications, the uses of the CRISPR system, and the past, present and future of CRISPR genome editing. Topics of interest in this updated volume include a section on CRISPR history, The genome editing revolution, Programming CRISPR and its applications, CRISPR Delivery methods, CRISPR libraries and screening, CRISPR investigation in haploid cells, CRISPR in the generation of transgenic animals, CRISPR therapeutics, and Promising strategies and present challenges. Accessible to students and researchers alike Written by leading authorities in the field
