15,060 research outputs found
Using the CRISPR/Cas9 system to understand neuropeptide biology and regulation
Funding was provided by a Wellcome Trust ISSF starting grant (105625/Z/14/Z), Medical Research Scotland (PhD-719-2013), GW Pharmaceuticals (PhD-719-2013 - S.5242.001) and the BBSRC (BB/J012343/1).Peer reviewedPublisher PD
Modulating signaling networks by CRISPR/Cas9-mediated transposable element insertion
In a recent past, transposable elements (TEs) were referred to as selfish genetic components only capable of copying themselves with the aim of increasing the odds of being inherited. Nonetheless, TEs have been initially proposed as positive control elements acting in synergy with the host. Nowadays, it is well known that TE movement into host genome comprises an important evolutionary mechanism capable of increasing the adaptive fitness. As insights into TE functioning are increasing day to day, the manipulation of transposition has raised an interesting possibility of setting the host functions, although the lack of appropriate genome engineering tools has unpaved it. Fortunately, the emergence of genome editing technologies based on programmable nucleases, and especially the arrival of a multipurpose RNA-guided Cas9 endonuclease system, has made it possible to reconsider this challenge. For such purpose, a particular type of transposons referred to as miniature inverted-repeat transposable elements (MITEs) has shown a series of interesting characteristics for designing functional drivers. Here, recent insights into MITE elements and versatile RNA-guided CRISPR/Cas9 genome engineering system are given to understand how to deploy the potential of TEs for control of the host transcriptional activity.Fil: Vaschetto, Luis Maria Benjamin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Cátedra de Diversidad Animal I; Argentin
Genome editing technologies to fight infectious diseases
Genome editing by programmable nucleases represents a promising tool that could be exploited to develop new therapeutic strategies to fight infectious diseases. These nucleases, such as zinc-finger nucleases, transcription activator-like effector nucleases, clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated protein 9 (Cas9) and homing endonucleases, are molecular scissors that can be targeted at predetermined loci in order to modify the genome sequence of an organism. Areas covered: By perturbing genomic DNA at predetermined loci, programmable nucleases can be used as antiviral and antimicrobial treatment. This approach includes targeting of essential viral genes or viral sequences able, once mutated, to inhibit viral replication; repurposing of CRISPR-Cas9 system for lethal self-targeting of bacteria; targeting antibiotic-resistance and virulence genes in bacteria, fungi, and parasites; engineering arthropod vectors to prevent vector-borne infections. Expert commentary: While progress has been done in demonstrating the feasibility of using genome editing as antimicrobial strategy, there are still many hurdles to overcome, such as the risk of off-target mutations, the raising of escape mutants, and the inefficiency of delivery methods, before translating results from preclinical studies into clinical applications
CRISPR/Cas9‐mediated genome editing: from basic research to translational medicine
The recent development of the CRISPR/Cas9 system as an efficient and accessible programmable genome-editing tool has revolutionized basic science research. CRISPR/Cas9 system-based technologies have armed researchers with new powerful tools to unveil the impact of genetics on disease development by enabling the creation of precise cellular and animal models of human diseases. The therapeutic potential of these technologies is tremendous, particularly in gene therapy, in which a patient-specific mutation is genetically corrected in order to treat human diseases that are untreatable with conventional therapies. However, the translation of CRISPR/Cas9 into the clinics will be challenging, since we still need to improve the efficiency, specificity and delivery of this technology. In this review, we focus on several in vitro, in vivo and ex vivo applications of the CRISPR/Cas9 system in human disease-focused research, explore the potential of this technology in translational medicine and discuss some of the major challenges for its future use in patients.Portuguese Foundation for Science and Technology:
UID/BIM/04773/2013
1334
Spanish Ministry of Science, Innovation and Universities
RTI2018-094629-B-I00
Portuguese Foundation for Science and Technology
SFRH/BPD/100434/2014
European Union (EU)
748585
LPCC-NRS/Terry Fox grantsinfo:eu-repo/semantics/publishedVersio
Developments in the tools and methodologies of synthetic biology.
Synthetic biology is principally concerned with the rational design and engineering of biologically based parts, devices, or systems. However, biological systems are generally complex and unpredictable, and are therefore, intrinsically difficult to engineer. In order to address these fundamental challenges, synthetic biology is aiming to unify a body of knowledge from several foundational scientific fields, within the context of a set of engineering principles. This shift in perspective is enabling synthetic biologists to address complexity, such that robust biological systems can be designed, assembled, and tested as part of a biological design cycle. The design cycle takes a forward-design approach in which a biological system is specified, modeled, analyzed, assembled, and its functionality tested. At each stage of the design cycle, an expanding repertoire of tools is being developed. In this review, we highlight several of these tools in terms of their applications and benefits to the synthetic biology community
Short Synthetic Terminators for Assembly of Transcription Units in vitro and Stable Chromosomal Integration in Yeast S. cerevisiae
The authors thank the assistance of the Iain Fraser Cytometry Centre at the University of Aberdeen. We also thank Claire MacGregor, Diane Massie and Yvonne Turnbull for technical assistance, Alexander Lorenz and Ryohei Sekido for critical reading of the manuscript and Richard Newton for preliminary results. This work was supported by Scottish Universities Life Sciences Alliance (SULSA).Peer reviewedPostprin
The physicist's guide to one of biotechnology's hottest new topics: CRISPR-Cas
Clustered regularly interspaced short palindromic repeats (CRISPR) and
CRISPR-associated proteins (Cas) constitute a multi-functional, constantly
evolving immune system in bacteria and archaea cells. A heritable, molecular
memory is generated of phage, plasmids, or other mobile genetic elements that
attempt to attack the cell. This memory is used to recognize and interfere with
subsequent invasions from the same genetic elements. This versatile prokaryotic
tool has also been used to advance applications in biotechnology. Here we
review a large body of CRISPR-Cas research to explore themes of evolution and
selection, population dynamics, horizontal gene transfer, specific and
cross-reactive interactions, cost and regulation, non-immunological CRISPR
functions that boost host cell robustness, as well as applicable mechanisms for
efficient and specific genetic engineering. We offer future directions that can
be addressed by the physics community. Physical understanding of the CRISPR-Cas
system will advance uses in biotechnology, such as developing cell lines and
animal models, cell labeling and information storage, combatting antibiotic
resistance, and human therapeutics.Comment: 75 pages, 15 figures, Physical Biology (2018
Physical Model of the Immune Response of Bacteria Against Bacteriophage Through the Adaptive CRISPR-Cas Immune System
Bacteria and archaea have evolved an adaptive, heritable immune system that
recognizes and protects against viruses or plasmids. This system, known as the
CRISPR-Cas system, allows the host to recognize and incorporate short foreign
DNA or RNA sequences, called `spacers' into its CRISPR system. Spacers in the
CRISPR system provide a record of the history of bacteria and phage
coevolution. We use a physical model to study the dynamics of this coevolution
as it evolves stochastically over time. We focus on the impact of mutation and
recombination on bacteria and phage evolution and evasion. We discuss the
effect of different spacer deletion mechanisms on the coevolutionary dynamics.
We make predictions about bacteria and phage population growth, spacer
diversity within the CRISPR locus, and spacer protection against the phage
population.Comment: 37 pages, 13 figure
- …