Emergence Of Genome Editing Technologies Research Articles

Revolutionizing Fish Biotechnology: A Current Status and Future Prospects

Fish biotechnology has witnessed significant advancements in recent years, driven by a convergence of interdisciplinary research efforts spanning genetics, genomics, aquaculture, and molecular biology. This synthesis study examines the significant advancements in fish biotechnology and presents potential opportunities for transforming the sector in the future. The emergence of genome editing technologies, such as CRISPR-Cas9, has fundamentally transformed the accurate manipulation of fish genomes, allowing for specific alterations in features that span from disease resistance to growth improvement. These methods show great potential for promoting sustainable aquaculture operations and protecting endangered species. In addition, the incorporation of omics technologies, like as genomics, transcriptomics, and proteomics, has yielded unparalleled understanding of the molecular pathways that underlie complex features in fish. This comprehensive approach has enabled the identification of crucial genes and regulatory circuits that control different physiological processes, therefore aiding the creation of new therapies for disease management and selective breeding. The recent advancements in fish biotechnology have had a significant and positive effect on the industry. These achievements highlight the potential for transforming the sector through collaboration across many disciplines, technical innovation, and the adoption of sustainable methods.

NTLA-2002: CRISPR/Cas9-mediated gene knockout of KLKB1 to treat hereditary angioedema

Hereditary angioedema (HAE) is a rare genetic disorder characterized by recurrent episodes of debilitating and potentially fatal swelling in various parts of the body including face, hand and airways. Prophylactic treatment options for HAE patients have dramatically improved in recent years, significantly reducing the frequency of attacks and overall disease burden but all require chronic, potentially life-long administration. The emergence of genome editing technologies as a new therapeutic modality offers the promise of curing many genetic diseases. To this end, Intellia Therapeutics is developing NTLA-2002, an investigational CRISPR/Cas9-based therapy targeting KLKB1 for the treatment of HAE. Guide RNA targeting human- and cynomolgus monkey-specific KLKB1 were identified and formulated using Intellia’s modular CRISPR/Cas9 lipid nanoparticle (LNP) platform technology. These formulations were evaluated in a humanized KLKB1 mouse model (huKLKB1) and the cynomolgus monkey. KLKB1 gene editing, plasma kallikrein concentration and activity, and vascular leakage were evaluated. In the huKLKB1 mouse, a single administration of NTLA-2002 resulted in robust KLKB1 gene editing (∼70%), subsequent reductions in total plasma kallikrein (>90%) and abrogation of captopril-induced vascular leakage. In the monkey, a single administration of cyno-specific LNP formulation resulted in robust gene editing (∼70%) and reductions in both total kallikrein protein and activity (>95%). Further, these reductions have been maintained for at least 15 months in an ongoing monkey study. A single administration of NTLA-2002 resulted in robust, durable reduction of kallikrein protein and activity, supporting further development as a potential one-time treatment option for patients with HAE.

Regulostat Inferelator: a novel network biology platform to uncover molecular devices that predetermine cellular response phenotypes.

With the emergence of genome editing technologies and synthetic biology, it is now possible to engineer genetic circuits driving a cell's phenotypic response to a stressor. However, capturing a continuous response, rather than simply a binary ‘on’ or ‘off’ response, remains a bioengineering challenge. No tools currently exist to identify gene candidates responsible for predetermining and fine-tuning cell response phenotypes. To address this gap, we devised a novel Regulostat Inferelator (RSI) algorithm to decipher intrinsic molecular devices or networks that predetermine cellular phenotypic responses. The RSI algorithm is designed to extract gene expression patterns from basal transcriptomic data in order to identify ‘regulostat’ constituent gene pairs, which exhibit rheostat-like mode-of-cooperation capable of fine-tuning cellular response. Our proof-of-concept study provides computational evidence for the existence of regulostats and that these networks predetermine cellular response prior to exposure to a stressor or drug. In addition, our work, for the first time, provides evidence of context-specific, drug–regulostat interactions in predetermining drug response phenotypes in cancer cells. Given RSI-inferred regulostat networks offer insights for prioritizing gene candidates capable of rendering a resistant phenotype sensitive to a given drug, we envision that this tool will be of great value in bioengineering and medicine.

Genome Editing with CRISPR/Cas9: First Steps Towards a new Era inMedicine?

The emergence of genome editing technologies can be regarded as one of the most groundbreaking revolutions in the history of science. Modern genome editing allows the introduction of precise mutations into the genome of virtually all cells and organisms without leaving any additional trace. Undoubtedly, genome editing with CRISPR/Cas9, often casually referred to as "genetic scissors", will revolutionize medical research and development. However, at the same time it creates a great need for ethical considerations as it might hold risks for both people and the environment that cannot yet be fully assessed. While genome editing is already well established in laboratory research, clinical applications based on genome editing are close. For the first time, targeted corrections of genetic defects in somatic cells, stem cells as well as in the germ line appear technically feasible. This generates possible future scenarios that urgently require broad ethical and social discussions.

Recent Progress in Genome Editing Approaches for Inherited Cardiovascular Diseases.

This review describes the recent progress in nuclease-based therapeutic applications for inherited heart diseases in vitro, highlights the development of the most recent genome editing technologies and discusses the associated challenges for clinical translation. Inherited cardiovascular disorders are passed from generation to generation. Over the past decade, considerable progress has been made in understanding the genetic basis of inherited heart diseases. The timely emergence of genome editing technologies using engineered programmable nucleases has revolutionized the basic research of inherited cardiovascular diseases and holds great promise for the development of targeted therapies. The genome editing toolbox is rapidly expanding, and new tools have been recently added that significantly expand the capabilities of engineered nucleases. Newer classes of versatile engineered nucleases, such as the "base editors," have been recently developed, offering the potential for efficient and precise therapeutic manipulation of the human genome.

Genome Editing in Human Neural Stem and Progenitor Cells.

Experimental tools for precise manipulation of mammalian genomes enable reverse genetic approaches to explore biology and disease. Powerful genome editing technologies built upon designer nucleases, such as CRISPR/Cas9, have recently emerged. Parallel progress has been made in methodologies for the expansion and differentiation of human pluripotent and tissue stem cells. Together these innovations provide a remarkable new toolbox for human cellular genetics and are opening up vast opportunities for discoveries and applications across the breadth of life sciences research. In this chapter, we review the emergence of genome editing technologies and how these are being deployed in studies of human neurobiology, neurological disease, and neuro-oncology. We focus our discussion on CRISPR/Cas9 and its application in studies of human neural stem and progenitor cells.

Application of genome editing technologies in rats for human disease models.

Laboratory rats and mice are representative experimental animals for models of human disease. The emergence of genome editing technologies has enabled us to produce a variety of genetically modified animals, including rats, as a means of elucidating the in vivo functions of the gene of interest and characterizing the molecular mechanisms of human disease. Several advanced techniques for knock-in methodologies in rats are currently in development, which permit researchers to introduce precise nucleotide modifications at target sites in the rat's genome. Furthermore, recent studies with knock-out rats have revealed that observed disease phenotypes are often more similar than mouse models to those of humans. In this article, we introduce the methodologies for efficient gene manipulation in rats using genome editing technologies, and describe the advances made using rats for human disease models. We also discuss the importance of gene manipulation in animal models for the better understanding of fundamental processes among different species.

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.

Genetic analysis of zebrafish gonadotropin (FSH and LH) functions by TALEN-mediated gene disruption.

Vertebrate reproduction is controlled by two gonadotropins (FSH and LH) from the pituitary. Despite numerous studies on FSH and LH in fish species, their functions in reproduction still remain poorly defined. This is partly due to the lack of powerful genetic approaches for functional studies in adult fish. This situation is now changing with the emergence of genome-editing technologies, especially Transcription Activator-Like Effector Nuclease (TALEN) and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR). In this study, we deleted the hormone-specific β-genes of both FSH and LH in the zebrafish using TALEN. This was followed by a phenotype analysis for key reproductive events, including gonadal differentiation, puberty onset, gametogenesis, final maturation, and fertility. FSH-deficient zebrafish (fshb(-/-)) were surprisingly fertile in both sexes; however, the development of both the ovary and testis was significantly delayed. In contrast, LH-deficient zebrafish (lhb(-/-)) showed normal gonadal growth, but the females failed to spawn and were therefore infertile. Using previtellogenic follicles as the marker, we observed a significant delay of puberty onset in the fshb mutant but not the lhb mutant females. Interestingly, FSH seemed to play a role in maintaining the female status because we repeatedly observed sexual reversal in the fshb mutant. Neither the fshb nor lhb mutation alone seemed to affect gonadal differentiation; however, the double mutation of the two genes led to all males, although the development of the testis was significantly delayed. In summary, our data confirmed some well-known functions of FSH and LH in fish while also providing evidence for novel functions, which would be difficult to reveal using traditional biochemical and physiological approaches.