Genome Editing in Zebrafish Using High-Fidelity Cas9 Nucleases: Choosing the Right Nuclease for the Task

Prykhozhij, Sergey V.; Cordeiro, Anna; Cáceres, Lucía; Berman, Jason N.

doi:10.1007/978-1-0716-0290-4_21

Cited by 6 publications

(4 citation statements)

References 31 publications

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“…Recent advances have expanded the range of available CRISPR nucleases, guide RNA design tools and other emerging technologies available to edit the zebrafish genome (Prykhozhij et al, 2018a;Liu et al, 2019). Off-target gene editing is a frequent problem when using CRISPR nucleases and this issue has been recently addressed both by the development of more sophisticated guide RNA design tools (Prykhozhij et al, 2018a;Liu et al, 2019) as well as by generating zebrafishoptimized vectors encoding HiFi Cas9 (Vakulskas et al, 2018) and highly specific but less efficient hypaCas9 (Chen et al, 2017) and testing them as mRNAs as well as commercial wild-type and HiFi Cas9 proteins using the same strategies (Prykhozhij et al, 2020).…”

Section: Genome Engineering Strategies For Cancer Predisposition Genetic Models In Zebrafishmentioning

confidence: 99%

Zebrafish Cancer Predisposition Models

Kobar

Collett

Prykhozhij

et al. 2021

Front. Cell Dev. Biol.

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Cancer predisposition syndromes are rare, typically monogenic disorders that result from germline mutations that increase the likelihood of developing cancer. Although these disorders are individually rare, resulting cancers collectively represent 5–10% of all malignancies. In addition to a greater incidence of cancer, affected individuals have an earlier tumor onset and are frequently subjected to long-term multi-modal cancer screening protocols for earlier detection and initiation of treatment. In vivo models are needed to better understand tumor-driving mechanisms, tailor patient screening approaches and develop targeted therapies to improve patient care and disease prognosis. The zebrafish (Danio rerio) has emerged as a robust model for cancer research due to its high fecundity, time- and cost-efficient genetic manipulation and real-time high-resolution imaging. Tumors developing in zebrafish cancer models are histologically and molecularly similar to their human counterparts, confirming the validity of these models. The zebrafish platform supports both large-scale random mutagenesis screens to identify potential candidate/modifier genes and recently optimized genome editing strategies. These techniques have greatly increased our ability to investigate the impact of certain mutations and how these lesions impact tumorigenesis and disease phenotype. These unique characteristics position the zebrafish as a powerful in vivo tool to model cancer predisposition syndromes and as such, several have already been created, including those recapitulating Li-Fraumeni syndrome, familial adenomatous polyposis, RASopathies, inherited bone marrow failure syndromes, and several other pathogenic mutations in cancer predisposition genes. In addition, the zebrafish platform supports medium- to high-throughput preclinical drug screening to identify compounds that may represent novel treatment paradigms or even prevent cancer evolution. This review will highlight and synthesize the findings from zebrafish cancer predisposition models created to date. We will discuss emerging trends in how these zebrafish cancer models can improve our understanding of the genetic mechanisms driving cancer predisposition and their potential to discover therapeutic and/or preventative compounds that change the natural history of disease for these vulnerable children, youth and adults.

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Section: Genome Engineering Strategies For Cancer Predisposition Genetic Models In Zebrafishmentioning

confidence: 99%

Zebrafish Cancer Predisposition Models

Kobar

Collett

Prykhozhij

et al. 2021

Front. Cell Dev. Biol.

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“…A protocol for the knockout of genes in zebrafish using HypaCas9 and HiFi Cas9 has been developed [ 207 ]. Scientists combined the SV40 NLS cell nucleus signaling protein with Cas12a for applications in zebrafish individuals [ 208 ].…”

Section: Genome Editing In Salmonidae and Cyprinidae Aquaculture Fish...mentioning

confidence: 99%

In Search of a Target Gene for a Desirable Phenotype in Aquaculture: Genome Editing of Cyprinidae and Salmonidae Species

Orlova,

Ruzina,

Emelianova

et al. 2024

Genes

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Aquaculture supplies the world food market with a significant amount of valuable protein. Highly productive aquaculture fishes can be derived by utilizing genome-editing methods, and the main problem is to choose a target gene to obtain the desirable phenotype. This paper presents a review of the studies of genome editing for genes controlling body development, growth, pigmentation and sex determination in five key aquaculture Salmonidae and Cyprinidae species, such as rainbow trout (Onchorhynchus mykiss), Atlantic salmon (Salmo salar), common carp (Cyprinus carpio), goldfish (Carassius auratus), Gibel carp (Carassius gibelio) and the model fish zebrafish (Danio rerio). Among the genes studied, the most applicable for aquaculture are mstnba, pomc, and acvr2, the knockout of which leads to enhanced muscle growth; runx2b, mutants of which do not form bones in myoseptae; lepr, whose lack of function makes fish fast-growing; fads2, Δ6abc/5Mt, and Δ6bcMt, affecting the composition of fatty acids in fish meat; dnd mettl3, and wnt4a, mutants of which are sterile; and disease-susceptibility genes prmt7, gab3, gcJAM-A, and cxcr3.2. Schemes for obtaining common carp populations consisting of only large females are promising for use in aquaculture. The immobilized and uncolored zebrafish line is of interest for laboratory use.

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“…Thus, when raising an F1 generation, each individual may have varying mutations and detailed analysis is necessary when finding stable lines. There have been continuing advances that reduce off-target modifications to the genome when employing CRISPR/Cas9, such as the development of high-fidelity Cas9 derivatives [ 131 ].…”

Section: Molecular Genetic Toolkit In the Zebrafish Animal Modelmentioning

confidence: 99%

Principles of Zebrafish Nephron Segment Development

Nguyen

Petrikas

Chambers

et al. 2023

JDB

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Nephrons are the functional units which comprise the kidney. Each nephron contains a number of physiologically unique populations of specialized epithelial cells that are organized into discrete domains known as segments. The principles of nephron segment development have been the subject of many studies in recent years. Understanding the mechanisms of nephrogenesis has enormous potential to expand our knowledge about the basis of congenital anomalies of the kidney and urinary tract (CAKUT), and to contribute to ongoing regenerative medicine efforts aimed at identifying renal repair mechanisms and generating replacement kidney tissue. The study of the zebrafish embryonic kidney, or pronephros, provides many opportunities to identify the genes and signaling pathways that control nephron segment development. Here, we describe recent advances of nephron segment patterning and differentiation in the zebrafish, with a focus on distal segment formation.

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Genome Editing in Zebrafish Using High-Fidelity Cas9 Nucleases: Choosing the Right Nuclease for the Task

Cited by 6 publications

References 31 publications

Zebrafish Cancer Predisposition Models

Zebrafish Cancer Predisposition Models

In Search of a Target Gene for a Desirable Phenotype in Aquaculture: Genome Editing of Cyprinidae and Salmonidae Species

Principles of Zebrafish Nephron Segment Development

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