From Gene Targeting to Genome Editing: Transgenic animals applications and beyond

ABSTRACTGenome modification technologies are powerful tools for molecular biology and related areas. Advances in animal transgenesis and genome editing technologies during the past three decades allowed systematic interrogation of gene function that can help model how the genome influences cellular physiology. Genetic engineering via homologous recombination (HR) has been the standard method to modify genomic sequences. Nevertheless, nuclease-guided genome editing methods that were developed recently, such as ZFN, TALEN and CRISPR/Cas, opened new perspectives for biomedical research. Here, we present a brief historical perspective of genome modification methods, focusing on transgenic mice models. Moreover, we describe how new techniques were discovered and improved, present the paradigm shifts and discuss their limitations and applications for biomedical research as well as possible future directions.

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Use of Customizable Nucleases for Gene Editing and Other Novel Applications

Genes ◽

10.3390/genes11090976 ◽

2020 ◽

Vol 11 (9) ◽

pp. 976

Author(s):

Pradeep Reddy ◽

Felipe Vilella ◽

Juan Carlos Izpisua Belmonte ◽

Carlos Simón

Keyword(s):

Animal Models ◽

Genome Editing ◽

Biomedical Research ◽

Laboratory Animal ◽

Gene Editing ◽

Genetic Disorders ◽

Basic Research ◽

The Past ◽

Novel Applications ◽

In Cells

The development of novel genome editing tools has unlocked new opportunities that were not previously possible in basic and biomedical research. During the last two decades, several new genome editing methods have been developed that can be customized to modify specific regions of the genome. However, in the past couple of years, many newer and more exciting genome editing techniques have been developed that are more efficient, precise, and easier to use. These genome editing tools have helped to improve our understanding of genetic disorders by modeling them in cells and animal models, in addition to correcting the disease-causing mutations. Among the genome editing tools, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system has proven to be the most popular one due to its versatility and has been successfully used in a wide variety of laboratory animal models and plants. In this review, we summarize the customizable nucleases currently used for genome editing and their uses beyond the modification of genome. We also discuss the potential future applications of gene editing tools for both basic research and clinical purposes.

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Paradigm shifts in electrophysiological mechanisms of atrial fibrillation

EP Europace ◽

10.1093/europace/euaa384 ◽

2020 ◽

Author(s):

Ulrich Schotten ◽

Seungyup Lee ◽

Stef Zeemering ◽

Albert L Waldo

Keyword(s):

Atrial Fibrillation ◽

Double Layers ◽

Paradigm Shifts ◽

New Techniques ◽

Activation Patterns ◽

Large Variability ◽

The Past ◽

Different Types ◽

Source Of Information ◽

Electrophysiological Mechanism

Abstract Determining the sequence of activation is a major source of information for understanding the electrophysiological mechanism(s) of atrial fibrillation (AF). However, the complex morphology of the electrograms hampers their analysis, and has stimulated generations of electrophysiologists to develop a large variety of technologies for recording, pre-processing, and analysis of fibrillation electrograms. This variability of approaches is mirrored by a large variability in the interpretation of fibrillation electrograms and, thereby, opinions regarding the basic electrophysiological mechanism(s) of AF vary widely. Multiple wavelets, different types of re-entry including rotors, double layers, multiple focal activation patterns all have been advocated, and a comprehensive and commonly accepted paradigm for the fundamental mechanisms of AF is still lacking. Here, we summarize the Maastricht perspective and Cleveland perspective regarding AF mechanism(s). We also describe some of the key observations in mapping of AF reported over the past decades, and how they changed over the years, often as results of new techniques introduced in the experimental field of AF research.

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Unlocking the Subconscious Consumer Bias: A Survey on the Past, Present, and Future of Hybrid EEG Schemes in Neuromarketing

Frontiers in Neuroergonomics ◽

10.3389/fnrgo.2021.672982 ◽

2021 ◽

Vol 2 ◽

Author(s):

Fotis P. Kalaganis ◽

Kostas Georgiadis ◽

Vangelis P. Oikonomou ◽

Nikos A. Laskaris ◽

Spiros Nikolopoulos ◽

...

Keyword(s):

Electrodermal Activity ◽

Brain Activity ◽

Relevant Literature ◽

Monitoring Methods ◽

Monitoring Method ◽

New Techniques ◽

Future Directions ◽

The Past ◽

Success Stories ◽

Holistic Approaches

Fueled by early success stories, the neuromarketing domain advanced rapidly during the last 10 years. As exciting new techniques were being adapted from medical research to the commercial domain, many neuroscientists and marketing practitioners have taken the chance to exploit them so as to uncover the answers of the most important marketing questions. Among the available neuroimaging technologies, electroencephalography (EEG) stands out as the less invasive and most affordable method. While not equally precise as other neuroimaging technologies in terms of spatial resolution, it can capture brain activity almost at the speed of cognition. Hence, EEG constitutes a favorable candidate for recording and subsequently decoding the consumers' brain activity. However, despite its wide use in neuromarketing, it cannot provide the complete picture alone. In order to overcome the limitations imposed by a single monitoring method, researchers focus on more holistic approaches. The exploitation of hybrid EEG schemes (e.g., combining EEG with eye-tracking, electrodermal activity, heart rate, and/or other) is ever growing and will hopefully allow neuromarketing to uncover consumers' behavior. Our survey revolves around last-decade hybrid neuromarketing schemes that involve EEG as the dominant modality. Beyond covering the relevant literature and state-of-the-art findings, we also provide future directions on the field, present the limitations that accompany each of the commonly employed monitoring methods and briefly discuss the omni-present ethical scepticizm related to neuromarketing.

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Stellar Evolution

Transactions of the International Astronomical Union ◽

10.1017/s0251107x00022343 ◽

1962 ◽

Vol 11 (02) ◽

pp. 137-143

Author(s):

M. Schwarzschild

Keyword(s):

Solar System ◽

Stellar Evolution ◽

Space Craft ◽

New Techniques ◽

Substantial Body ◽

Individual Stars ◽

The Past ◽

Evolution Of Galaxies ◽

History Of ◽

Fast Flow

It is perhaps one of the most important characteristics of the past decade in astronomy that the evolution of some major classes of astronomical objects has become accessible to detailed research. The theory of the evolution of individual stars has developed into a substantial body of quantitative investigations. The evolution of galaxies, particularly of our own, has clearly become a subject for serious research. Even the history of the solar system, this close-by intriguing puzzle, may soon make the transition from being a subject of speculation to being a subject of detailed study in view of the fast flow of new data obtained with new techniques, including space-craft.

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On Future Directions in Pathology

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100053991 ◽

1982 ◽

Vol 40 ◽

pp. 274-277

Author(s):

Benjamin F. Trump ◽

Irene K. Berezesky ◽

Raymond T. Jones

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Clinical Pathology ◽

Future Directions ◽

Diagnostic Pathology ◽

The Past ◽

Transmission Electron ◽

Organ Systems ◽

The Many

The role of electron microscopy and associated techniques is assured in diagnostic pathology. At the present time, most of the progress has been made on tissues examined by transmission electron microscopy (TEM) and correlated with light microscopy (LM) and by cytochemistry using both plastic and paraffin-embedded materials. As mentioned elsewhere in this symposium, this has revolutionized many fields of pathology including diagnostic, anatomic and clinical pathology. It began with the kidney; however, it has now been extended to most other organ systems and to tumor diagnosis in general. The results of the past few years tend to indicate the future directions and needs of this expanding field. Now, in addition to routine EM, pathologists have access to the many newly developed methods and instruments mentioned below which should aid considerably not only in diagnostic pathology but in investigative pathology as well.

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Ultrastructure and analytical microprobe analysis of simian lung mite pigments

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100143249 ◽

1986 ◽

Vol 44 ◽

pp. 324-325

Author(s):

R. W. Cole ◽

J. C. Kim

Keyword(s):

Biomedical Research ◽

Tissue Damage ◽

Microprobe Analysis ◽

Old World Monkeys ◽

Old World ◽

Experimental Animals ◽

The Past ◽

Lung Mite ◽

Mite Infestations ◽

Cardiopulmonary System

In recent years, non-human primates have become indispensable as experimental animals in many fields of biomedical research. Pharmaceutical and related industries alone use about 2000,000 primates a year. Respiratory mite infestations in lungs of old world monkeys are of particular concern because the resulting tissue damage can directly effect experimental results, especially in those studies involving the cardiopulmonary system. There has been increasing documentation of primate parasitology in the past twenty years.

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Comparison and Analysis of Computational Methods for Identifying N6 - methyladenosine Sites in Saccharomyces Cerevisiae

Current Pharmaceutical Design ◽

10.2174/1381612826666201109110703 ◽

2020 ◽

Vol 26 ◽

Author(s):

Pengmian Feng ◽

Lijing Feng ◽

Chaohui Tang

Keyword(s):

Saccharomyces Cerevisiae ◽

Computational Methods ◽

Computational Analysis ◽

Computational Prediction ◽

Experimental Methods ◽

Biological Processes ◽

Biological Functions ◽

Precise Location ◽

Future Directions ◽

The Past

Background and Purpose: N 6 -methyladenosine (m6A) plays critical roles in a broad set of biological processes. Knowledge about the precise location of m6A site in the transcriptome is vital for deciphering its biological functions. Although experimental techniques have made substantial contributions to identify m6A, they are still labor intensive and time consuming. As good complements to experimental methods, in the past few years, a series of computational approaches have been proposed to identify m6A sites. Methods: In order to facilitate researchers to select appropriate methods for identifying m6A sites, it is necessary to give a comprehensive review and comparison on existing methods. Results: Since researches on m6A in Saccharomyces cerevisiae are relatively clear, in this review, we summarized recent progresses on computational prediction of m6A sites in S. cerevisiae and assessed the performance of existing computational methods. Finally, future directions of computationally identifying m6A sites were presented. Conclusion: Taken together, we anticipate that this review will provide important guides for computational analysis of m 6A modifications.

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The Use of Autogenous Teeth for Alveolar Ridge Preservation: A Literature Review

Applied Sciences ◽

10.3390/app11041853 ◽

2021 ◽

Vol 11 (4) ◽

pp. 1853

Author(s):

João Cenicante ◽

João Botelho ◽

Vanessa Machado ◽

José João Mendes ◽

Paulo Mascarenhas ◽

...

Keyword(s):

Ridge Preservation ◽

Graft Material ◽

Alveolar Ridge ◽

Natural Consequence ◽

Oral Rehabilitation ◽

Future Directions ◽

The Past ◽

Alveolar Ridge Preservation ◽

Functional Consequences ◽

Teeth Extraction

Alveolar ridge resorption is a natural consequence of teeth extraction, with unpleasant aesthetic and functional consequences that might compromise a future oral rehabilitation. To minimize the biological consequences of alveolar ridge resorption, several surgical procedures have been designed, the so-called alveolar ridge preservation (ARP) techniques. One important characteristic is the concomitant use of biomaterial in ARP. In the past decade, autogenous teeth as a bone graft material in post-extraction sockets have been proposed with very interesting outcomes, yet with different protocols of preparation. Here we summarize the available evidence on autogenous teeth as a biomaterial in ARP, its different protocols and future directions.

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Small Molecule Inhibitors of Influenza Virus Entry

Pharmaceuticals ◽

10.3390/ph14060587 ◽

2021 ◽

Vol 14 (6) ◽

pp. 587

Author(s):

Zhaoyu Chen ◽

Qinghua Cui ◽

Michael Caffrey ◽

Lijun Rong ◽

Ruikun Du

Keyword(s):

Influenza Virus ◽

Membrane Fusion ◽

Small Molecule ◽

Drug Target ◽

Small Molecule Inhibitors ◽

Virus Entry ◽

Critical Role ◽

Structural Basis ◽

Future Directions ◽

The Past

Hemagglutinin (HA) plays a critical role during influenza virus receptor binding and subsequent membrane fusion process, thus HA has become a promising drug target. For the past several decades, we and other researchers have discovered a series of HA inhibitors mainly targeting its fusion machinery. In this review, we summarize the advances in HA-targeted development of small molecule inhibitors. Moreover, we discuss the structural basis and mode of action of these inhibitors, and speculate upon future directions toward more potent inhibitors of membrane fusion and potential anti-influenza drugs.

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The Past, Present and Future of Cannabis sativa Tissue Culture

Plants ◽

10.3390/plants10010185 ◽

2021 ◽

Vol 10 (1) ◽

pp. 185

Author(s):

Adrian S. Monthony ◽

Serena R. Page ◽

Mohsen Hesami ◽

Andrew Maxwell P. Jones

Keyword(s):

Tissue Culture ◽

Disease Virus ◽

Cannabis Sativa ◽

Clonal Plants ◽

Future Directions ◽

The Past ◽

Recent Developments ◽

History Of ◽

Drug Type

The recent legalization of Cannabis sativa L. in many regions has revealed a need for effective propagation and biotechnologies for the species. Micropropagation affords researchers and producers methods to rapidly propagate insect-/disease-/virus-free clonal plants and store germplasm and forms the basis for other biotechnologies. Despite this need, research in the area is limited due to the long history of prohibitions and restrictions. Existing literature has multiple limitations: many publications use hemp as a proxy for drug-type Cannabis when it is well established that there is significant genotype specificity; studies using drug-type cultivars are predominantly optimized using a single cultivar; most protocols have not been replicated by independent groups, and some attempts demonstrate a lack of reproducibility across genotypes. Due to culture decline and other problems, the multiplication phase of micropropagation (Stage 2) has not been fully developed in many reports. This review will provide a brief background on the history and botany of Cannabis as well as a comprehensive and critical summary of Cannabis tissue culture. Special attention will be paid to current challenges faced by researchers, the limitations of existing Cannabis micropropagation studies, and recent developments and future directions of Cannabis tissue culture technologies.

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