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Crispr-Cas

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https://www.readbyqxmd.com/read/29670305/germline-genetic-modification-and-identity-the-mitochondrial-and-nuclear-genomes
#1
COMMENT
Rosamund Scott, Stephen Wilkinson
In a legal 'first', the UK removed a prohibition against modifying embryos in human reproduction, to enable mitochondrial replacement techniques (MRTs), a move the Government distanced from 'germline genetic modification', which it aligned with modifying the nuclear genome. This paper (1) analyzes the uses and meanings of this term in UK/US legal and policy debates; and (2) evaluates related ethical concerns about identity. It shows that, with respect to identity, MRTs and nuclear genome editing techniques such as CRISPR/Cas-9 (now a policy topic), are not as different as has been supposed...
December 2017: Oxford Journal of Legal Studies
https://www.readbyqxmd.com/read/29668265/receptor-mediated-delivery-of-crispr-cas9-endonuclease-for-cell-type-specific-gene-editing
#2
Romain Rouet, Benjamin A Thuma, Marc D Roy, Nathanael G Lintner, David M Rubitski, James E Finley, Hanna M Wisniewska, Rima Mendonsa, Ariana Hirsh, Lorena de Oñate, Joan Compte, Thomas J Mclellan, Justin Bellenger, Xidong Feng, Alison Varghese, Boris A Chrunyk, Kris A Borzilleri, Kevin D Hesp, Kaihong Zhou, Nannan Ma, Meihua Tu, Robert Dullea, Kim F Mcclure, Ross C Wilson, Spiros Liras, Vincent Mascitti, Jennifer A Doudna
CRISPR-Cas RNA-guided endonucleases hold great promise for disrupting or correcting genomic sequences through site-specific DNA cleavage and repair. However, the lack of methods for cell- and tissue-selective delivery currently limits both research and clinical uses of these enzymes. We report the design and in vitro evaluation of S. pyogenes Cas9 proteins harboring asialoglycoprotein receptor ligands (ASGPrL). In particular, we demonstrate that the resulting ribonucleoproteins (Cas9-ASGPrL RNP) can be engineered to be preferentially internalized into cells expressing the corresponding receptor on their surface...
April 18, 2018: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/29668111/efficient-in-planta-gene-targeting-in-tomato-using-geminiviral-replicons-and-the-crispr-cas9-system
#3
Tal Dahan-Meir, Shdema Filler-Hayut, Cathy Melamed-Bessudo, Samuel Bocobza, Henryk Czosnek, Asaph Aharoni, Avraham A Levy
Current breeding relies mostly on random mutagenesis and recombination to generate novel genetic variation. However, targeted genome editing is becoming an increasingly important tool for precise plant breeding. Using the CRISPR-Cas system combined with the bean yellow dwarf virus rolling circle replicon we optimized a method for targeted mutagenesis and gene replacement in tomato. The carotenoid isomerase (CRTISO) and phytoene synthase 1 (PSY1) genes from the carotenoid biosynthesis pathway were chosen as targets due to their easily detectable change of phenotype...
April 18, 2018: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/29666786/modeling-neuropsychiatric-and-neurodegenerative-diseases-with-induced-pluripotent-stem-cells
#4
REVIEW
Elizabeth A LaMarca, Samuel K Powell, Schahram Akbarian, Kristen J Brennand
Human-induced pluripotent stem cells (hiPSCs) have revolutionized our ability to model neuropsychiatric and neurodegenerative diseases, and recent progress in the field is paving the way for improved therapeutics. In this review, we discuss major advances in generating hiPSC-derived neural cells and cutting-edge techniques that are transforming hiPSC technology, such as three-dimensional "mini-brains" and clustered, regularly interspersed short palindromic repeats (CRISPR)-Cas systems. We examine specific examples of how hiPSC-derived neural cells are being used to uncover the pathophysiology of schizophrenia and Parkinson's disease, and consider the future of this groundbreaking research...
2018: Frontiers in Pediatrics
https://www.readbyqxmd.com/read/29666291/determining-the-specificity-of-cascade-binding-interference-and-primed-adaptation-in-vivo-in-the-escherichia-coli-type-i-e-crispr-cas-system
#5
Lauren A Cooper, Anne M Stringer, Joseph T Wade
In clustered regularly interspaced short palindromic repeat (CRISPR)-Cas (CRISPR-associated) immunity systems, short CRISPR RNAs (crRNAs) are bound by Cas proteins, and these complexes target invading nucleic acid molecules for degradation in a process known as interference. In type I CRISPR-Cas systems, the Cas protein complex that binds DNA is known as Cascade. Association of Cascade with target DNA can also lead to acquisition of new immunity elements in a process known as primed adaptation. Here, we assess the specificity determinants for Cascade-DNA interaction, interference, and primed adaptation in vivo , for the type I-E system of Escherichia coli Remarkably, as few as 5 bp of crRNA-DNA are sufficient for association of Cascade with a DNA target...
April 17, 2018: MBio
https://www.readbyqxmd.com/read/29663663/modular-ligation-extension-of-guide-rna-operons-lego-for-multiplexed-dcas9-regulation-of-metabolic-pathways-in-saccharomyces-cerevisiae
#6
Matthew Deaner, Allison Holzman, Hal S Alper
Metabolic engineering typically utilizes a suboptimal step-wise gene target optimization approach to parse a highly connected and regulated cellular metabolism. While the endonuclease-null CRISPR/Cas system has enabled gene expression perturbations without genetic modification, it has been mostly limited to small sets of gene targets in eukaryotes due to inefficient methods to assemble and express large sgRNA operons. In this work, we develop a TEF1p-tRNA expression system and demonstrate that the use of tRNAs as splicing elements flanking sgRNAs provides higher efficiency than both Pol III and ribozyme-based expression across a variety of single sgRNA and multiplexed contexts...
April 16, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29661863/-vibrio-cholerae-outer-membrane-vesicles-inhibit-bacteriophage-infection
#7
Tamara Reyes-Robles, Rebecca S Dillard, Lynne S Cairns, Cecilia A Silva-Valenzuela, Max Housman, Afsar Ali, Elizabeth R Wright, Andrew Camilli
Novel preventatives could help in efforts to limit Vibrio cholerae infection and the spread of cholera. Bacteriophage (or phage) treatment has been proposed to be an alternative intervention, given the rapid replication of virulent phages, prey specificity, and relative ease of finding new virulent phages. Phage tropism is dictated in part by the presence of phage receptors on the bacterial surface. While many phages that can kill V. cholerae have been isolated, whether this pathogen is able to defend itself by neutralizing phage binding is unknown...
April 16, 2018: Journal of Bacteriology
https://www.readbyqxmd.com/read/29658880/crystal-structure-of-intraflagellar-transport-protein-80-reveals-a-homo-dimer-required-for-ciliogenesis
#8
Michael Taschner, Anna Lorentzen, André Mourão, Toby Collins, Grace M Freke, Dale Moulding, Jerome Basquin, Dagan Jenkins, Esben Lorentzen
Oligomeric assemblies of intraflagellar transport (IFT) particles build cilia through sequential recruitment and transport of ciliary cargo proteins within cilia. Here we present the 1.8Å resolution crystal structure of the Chlamydomonas IFT-B protein IFT80, which reveals the architecture of two N-terminal b-propellers followed by an a-helical extension. The N-terminal b-propeller tethers IFT80 to the IFT-B complex via IFT38 whereas the second b-propeller and the C-terminal a-helical extension result in IFT80 homo-dimerization...
April 16, 2018: ELife
https://www.readbyqxmd.com/read/29653191/microbiota-transplantation-and-or-crispr-cas-in-the-battle-against-antimicrobial-resistance
#9
EDITORIAL
Jordi Vila
No abstract text is available yet for this article.
April 10, 2018: Clinical Microbiology and Infection
https://www.readbyqxmd.com/read/29649958/the-nuts-and-bolts-of-the-haloferax-crispr-cas-system-i-b
#10
Lisa-Katharina Maier, Aris-Edda Stachler, Jutta Brendel, Britta Stoll, Susan Fischer, Karina Haas, Thandi Schwarz, Omer S Alkhnbashi, Kundan Sharma, Henning Urlaub, Rolf Backofen, Uri Gophna, Anita Marchfelder
Invading genetic elements pose a constant threat to prokaryotic survival, requiring an effective defence. Eleven years ago, the arsenal of known defence mechanisms was expanded by the discovery of the CRISPR-Cas system. Although CRISPR-Cas is present in the majority of archaea, research often focuses on bacterial models. Here, we provide a perspective based on insights gained studying CRISPR-Cas system I-B of the archaeon Haloferax volcanii. The system relies on more than 50 different crRNAs, whose stability and maintenance critically depend on the proteins Cas5 and Cas7, which bind the crRNA and form the Cascade complex...
April 13, 2018: RNA Biology
https://www.readbyqxmd.com/read/29648539/horizontal-gene-transfer-in-thermus-spp
#11
Alba Blesa, Beate Averhoff, José Berenguer
The small amount of genetic content in thermophiles generally limits their adaptability to environmental changes. In Thermus spp., very active horizontal gene transfer (HGT) mechanisms allow the rapid spread of strain-specific adaptive gene modules among the entire population. Constitutive expression of a rather particular and highly efficient DNA transport apparatus (DTA) is at the center of this HGT-mediated enhanced adaptability. The function of the DTA is dependent on the integrity and longevity of the extracellular DNA (eDNA) being transformed, which can be improved by the production of extracellular vesicles (EV) through lysis of a fraction of the population...
April 12, 2018: Current Issues in Molecular Biology
https://www.readbyqxmd.com/read/29644494/review-of-crispr-cas9-sgrna-design-tools
#12
REVIEW
Yingbo Cui, Jiaming Xu, Minxia Cheng, Xiangke Liao, Shaoliang Peng
The adaptive immunity system in bacteria and archaea, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-associate (CRISPR/Cas), has been adapted as a powerful gene editing tool and got a broad application in genome research field due to its ease of use and cost-effectiveness. The performance of CRISPR/Cas relies on well-designed single-guide RNA (sgRNA), so a lot of bioinformatic tools have been developed to assist the design of highly active and specific sgRNA. These tools vary in design specifications, parameters, genomes and so on...
April 11, 2018: Interdisciplinary Sciences, Computational Life Sciences
https://www.readbyqxmd.com/read/29642627/a-review-of-the-interactions-between-wheat-and-wheat-pathogens-zymoseptoria-tritici-fusarium-spp-and-parastagonospora-nodorum
#13
REVIEW
Adrian Duba, Klaudia Goriewa-Duba, Urszula Wachowska
Zymoseptoria tritici is a hemibiotrophic pathogen which causes Septoria leaf blotch in wheat. The pathogenesis of the disease consists of a biotrophic phase and a necrotrophic phase. The pathogen infects the host plant by suppressing its immune response in the first stage of infection. Hemibiotrophic pathogens of the genus Fusarium cause Fusarium head blight, and the necrotrophic Parastagonospora nodorum is responsible for Septoria nodorum blotch in wheat. Cell wall-degrading enzymes in plants promote infections by necrotrophic and hemibiotrophic pathogens, and trichothecenes, secondary fungal metabolites, facilitate infections caused by fungi of the genus Fusarium ...
April 10, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29629622/tolerance-of-sulfolobus-smv1-virus-to-the-immunity-of-i-a-and-iii-b-crispr-cas-systems-in-sulfolobus-islandicus
#14
Tong Guo, Wenyuan Han, Qunxin She
Sulfolobus islandicus Rey15A encodes one type I-A and two type III-B systems, all of which are active in mediating nucleic acids interference. However, the effectiveness of each CRISPR system against virus infection was not tested in this archaeon. Here we constructed S. islandicus strains that constitutively express the antiviral immunity from either I-A, or III-B, or I-A plus III-B systems against SMV1 and tested the response of each host to SMV1 infection. We found that, although both CRISPR immunities showed a strongly inhibition to viral DNA replication at an early stage of incubation, the host I-A CRISPR immunity gradually lost the control on virus proliferation, allowing accumulation of cellular viral DNA and release of a large number of viral particles...
April 9, 2018: RNA Biology
https://www.readbyqxmd.com/read/29621180/applications-of-crispr-cas-system-to-bacterial-metabolic-engineering
#15
REVIEW
Suhyung Cho, Jongoh Shin, Byung-Kwan Cho
The clustered regularly interspaced short palindromic repeats/CRISPR-associated (CRISPR/Cas) adaptive immune system has been extensively used for gene editing, including gene deletion, insertion, and replacement in bacterial and eukaryotic cells owing to its simple, rapid, and efficient activities in unprecedented resolution. Furthermore, the CRISPR interference (CRISPRi) system including deactivated Cas9 (dCas9) with inactivated endonuclease activity has been further investigated for regulation of the target gene transiently or constitutively, avoiding cell death by disruption of genome...
April 5, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29618800/a-rapid-crispr-cas-based-mutagenesis-assay-in-zebrafish-for-identification-of-genes-involved-in-thyroid-morphogenesis-and-function
#16
A Trubiroha, P Gillotay, N Giusti, D Gacquer, F Libert, A Lefort, B Haerlingen, X De Deken, R Opitz, S Costagliola
The foregut endoderm gives rise to several organs including liver, pancreas, lung and thyroid with important roles in human physiology. Understanding which genes and signalling pathways regulate their development is crucial for understanding developmental disorders as well as diseases in adulthood. We exploited unique advantages of the zebrafish model to develop a rapid and scalable CRISPR/Cas-based mutagenesis strategy aiming at the identification of genes involved in morphogenesis and function of the thyroid...
April 4, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29616920/correction-type-iii-crispr-cas-systems-can-provide-redundancy-to-counteract-viral-escape-from-type-i-systems
#17
Sukrit Silas, Patricia Lucas-Elio, Simon A Jackson, Alejandra Aroca-Crevillén, Loren L Hansen, Peter C Fineran, Andrew Z Fire, Antonio Sánchez-Amat
No abstract text is available yet for this article.
April 4, 2018: ELife
https://www.readbyqxmd.com/read/29616594/the-protective-function-of-noncoding-dna-in-genome-defense-of-eukaryotic-male-germ-cells
#18
Guo-Hua Qiu, Cuiqin Huang, Xintian Zheng, Xiaoyan Yang
Peripheral and abundant noncoding DNA has been hypothesized to protect the genome and the central protein-coding sequences against DNA damage in somatic genome. In the cytosol, invading exogenous nucleic acids may first be deactivated by small RNAs encoded by noncoding DNA via mechanisms similar to the prokaryotic CRISPR-Cas system. In the nucleus, the radicals generated by radiation in the cytosol, radiation energy and invading exogenous nucleic acids are absorbed, blocked and/or reduced by peripheral heterochromatin, and damaged DNA in heterochromatin is removed and excluded from the nucleus to the cytoplasm through nuclear pore complexes...
April 4, 2018: Epigenomics
https://www.readbyqxmd.com/read/29608720/homologous-recombination-between-genetically-divergent-campylobacter-fetus-lineages-supports-host-associated-speciation
#19
Maarten J Gilbert, Birgitta Duim, Linda van der Graaf-van Bloois, Jaap A Wagenaar, Aldert L Zomer
Homologous recombination is a major driver of bacterial speciation. Genetic divergence and host association are important factors influencing homologous recombination. Here, we study these factors for Campylobacter fetus, which shows a distinct intraspecific host dichotomy. Campylobacter fetus subspecies fetus (Cff) and venerealis are associated with mammals, whereas C. fetus subsp. testudinum (Cft) is associated with reptiles. Recombination between these genetically divergent C. fetus lineages is extremely rare...
March 1, 2018: Genome Biology and Evolution
https://www.readbyqxmd.com/read/29605859/functional-insulator-scanning-of-cpg-islands-to-identify-regulatory-regions-of-promoters-using-crispr
#20
Alice Grob, Masue Marbiah, Mark Isalan
The ability to mutate a promoter in situ is potentially a very useful approach for gaining insights into endogenous gene regulation mechanisms. The advent of CRISPR/Cas systems has provided simple, efficient, and targeted genetic manipulation in eukaryotes, which can be applied to studying genome structure and function.The basic CRISPR toolkit comprises an endonuclease, Cas9, and a short DNA-targeting sequence, made up of a single guide RNA (sgRNA). The catalytic domains of Cas9 are rendered active upon dimerization of Cas9 with sgRNA, resulting in targeted double stranded DNA breaks...
2018: Methods in Molecular Biology
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