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Transgenic and Genetically Engineered Animal and Cell Culture Models
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Transgenic and Genetically Engineered Animal and Cell Culture Models

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Genetics and Genomics".

Deadline for manuscript submissions: closed (15 November 2023) | Viewed by 6724

Special Issue Editors

Prof. Dr. Wilfried A. Kues

E-Mail Website
Guest Editor
Friedrich-Loeffler-Institute, Department of Biotechnology, Neustadt, Germany
Interests: epigenetic mechanisms of heredity; transgenesis; genetic engineering; synthetic biology; cell biology; developmental biology; stem cells
Prof. Dr. Goo Jang

E-Mail Website
Guest Editor
College of Veterinary Medicine, Seoul National University, Seoul, Korea
Interests: transgenesis; genetic engineering (gene editing); (farm) animal developmental biology

Special Issue Information

Dear Colleagues,

In recent years, seminal discoveries in the fields of whole-genome sequencing, genetic engineering, and cellular reprogramming accelerated the progress and methodological repertoire of gene transfer into vertebrate (and invertebrate) cells, oocytes, and embryos for the study of developmental, genetic, reproductive, and disease-related biological questions.

Since the “start point” of genetic engineering, the Asilomar Conference on Recombinant DNA in 1975, this research direction has made impressive progress in terms of speed, specificity, and efficiency, and has accumulated a vast amount of biological information and models for analyzing biological phenomena.

In the early years the eukaryotic (and even viral and prokaryotic) genomes were terra incognita, and researchers need laborious methods to identify, subclone, and characterize a particular gene. This changed with the development of whole-genome sequencing methods, accompanied by the identification and development of genetic engineering techniques based on the activity of ectopic enzymes, such as recombinases, transposases, and programmable nucleases. Similarly, endogenous mechanisms such as RNA editing and RNA interference were discovered and employed to address specific questions. Fluorescent proteins, with the prototypic enhanced green fluorescent protein (EGFP), became widespread tools and allowed the live tracking of individual cells. Genetically engineered cell and organoid cultures, but also transgenic and genome-edited organisms, were developed. For the implementation of these techniques in mammalian embryos, the development of sophisticated methods such as blastocyst complementation and somatic-cell nuclear transfer were essential. Further advancements include the single-cell sequencing of DNA, RNA and even single-cell proteomics. A spin out of these advancements is the field of synthetic biology, which aims to construct optimized pathways and even cells from scratch.

In summary, progress in basic research but also a flurry of applications, such as the production of safe recombinant proteins for industrial uses and therapeutic applications, transgenic animals with improved traits or which produce complex proteins, cellular reprogramming, and the first gene therapies for human patients have been realized. The rapid development of effective COVID-19 vaccines, based either on synthetic messenger RNA or on recombinant vectors, is just one prominent example.

This Special Issue aims to assemble a snapshot of the current state of the art in “gene transfer” in vertebrate cells and animals, but articles and reviews in neighboring fields are also welcome, since this field is dynamically developing and new directions are just emerging.

Prof. Dr. Wilfried A. Kues
Prof. Dr. Goo Jang
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • CRISPR/Cas
  • genome editing tools
  • recombinase
  • transposon
  • RNAi
  • EGFP
  • embryonic stem cells
  • induced pluripotent stem cells
  • regenerative medicine
  • epigenetics
  • RNA interference
  • RNA editing
  • transgenic animals
  • genetically engineered animals
  • synthetic biology

Published Papers (3 papers)

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Research

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11 pages, 1884 KiB  
Article
Pig Coat Color Manipulation by MC1R Gene Editing
by Haiwen Zhong, Jian Zhang, Cheng Tan, Junsong Shi, Jie Yang, Gengyuan Cai, Zhenfang Wu and Huaqiang Yang
Int. J. Mol. Sci. 2022, 23(18), 10356; https://doi.org/10.3390/ijms231810356 - 8 Sep 2022
Cited by 3 | Viewed by 2618
Abstract
Black coat color in pigs is determined by the dominant E allele at the MC1R locus. Through comparing MC1R gene sequences between recessive e and dominant ED1 alleles, we identified four missense mutations that could affect MC1R protein function for eumelanin synthesis. [...] Read more.
Black coat color in pigs is determined by the dominant E allele at the MC1R locus. Through comparing MC1R gene sequences between recessive e and dominant ED1 alleles, we identified four missense mutations that could affect MC1R protein function for eumelanin synthesis. With the aim of devising a genetic modification method for pig coat color manipulation, we mutated the e allele in the Duroc breed to the dominant ED1 allele using CRISPR-mediated homologous recombination for the four mutation substitutions at the MC1R locus. The MC1R-modified Duroc pigs generated using the allele replacement strategy displayed uniform black coat color across the body. A genotyping assay showed that the MC1R-modified Duroc pigs had a heterozygous ED1/e allele at the MC1R locus; in addition, the pigs remained in the Duroc genetic background. Our work offers a gene editing method for pig coat color manipulation, which could value the culture of new pig varieties meeting the needs of diversified market. Full article
(This article belongs to the Special Issue Transgenic and Genetically Engineered Animal and Cell Culture Models)
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Review

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21 pages, 2394 KiB  
Review
Animal Models for the Study of Gaucher Disease
by Or Cabasso, Aparna Kuppuramalingam, Lindsey Lelieveld, Martijn Van der Lienden, Rolf Boot, Johannes M. Aerts and Mia Horowitz
Int. J. Mol. Sci. 2023, 24(22), 16035; https://doi.org/10.3390/ijms242216035 - 7 Nov 2023
Cited by 2 | Viewed by 1307
Abstract
In Gaucher disease (GD), a relatively common sphingolipidosis, the mutant lysosomal enzyme acid β-glucocerebrosidase (GCase), encoded by the GBA1 gene, fails to properly hydrolyze the sphingolipid glucosylceramide (GlcCer) in lysosomes, particularly of tissue macrophages. As a result, GlcCer accumulates, which, to a certain [...] Read more.
In Gaucher disease (GD), a relatively common sphingolipidosis, the mutant lysosomal enzyme acid β-glucocerebrosidase (GCase), encoded by the GBA1 gene, fails to properly hydrolyze the sphingolipid glucosylceramide (GlcCer) in lysosomes, particularly of tissue macrophages. As a result, GlcCer accumulates, which, to a certain extent, is converted to its deacylated form, glucosylsphingosine (GlcSph), by lysosomal acid ceramidase. The inability of mutant GCase to degrade GlcSph further promotes its accumulation. The amount of mutant GCase in lysosomes depends on the amount of mutant ER enzyme that shuttles to them. In the case of many mutant GCase forms, the enzyme is largely misfolded in the ER. Only a fraction correctly folds and is subsequently trafficked to the lysosomes, while the rest of the misfolded mutant GCase protein undergoes ER-associated degradation (ERAD). The retention of misfolded mutant GCase in the ER induces ER stress, which evokes a stress response known as the unfolded protein response (UPR). GD is remarkably heterogeneous in clinical manifestation, including the variant without CNS involvement (type 1), and acute and subacute neuronopathic variants (types 2 and 3). The present review discusses animal models developed to study the molecular and cellular mechanisms underlying GD. Full article
(This article belongs to the Special Issue Transgenic and Genetically Engineered Animal and Cell Culture Models)
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23 pages, 1700 KiB  
Review
Genetically Engineered Mice Unveil In Vivo Roles of the Mediator Complex
by Leonid A. Ilchuk, Marina V. Kubekina, Yulia D. Okulova, Yulia Yu. Silaeva, Victor V. Tatarskiy, Maxim A. Filatov and Alexandra V. Bruter
Int. J. Mol. Sci. 2023, 24(11), 9330; https://doi.org/10.3390/ijms24119330 - 26 May 2023
Viewed by 1625
Abstract
The Mediator complex is a multi-subunit protein complex which plays a significant role in the regulation of eukaryotic gene transcription. It provides a platform for the interaction of transcriptional factors and RNA polymerase II, thus coupling external and internal stimuli with transcriptional programs. [...] Read more.
The Mediator complex is a multi-subunit protein complex which plays a significant role in the regulation of eukaryotic gene transcription. It provides a platform for the interaction of transcriptional factors and RNA polymerase II, thus coupling external and internal stimuli with transcriptional programs. Molecular mechanisms underlying Mediator functioning are intensively studied, although most often using simple models such as tumor cell lines and yeast. Transgenic mouse models are required to study the role of Mediator components in physiological processes, disease, and development. As constitutive knockouts of most of the Mediator protein coding genes are embryonically lethal, conditional knockouts and corresponding activator strains are needed for these studies. Recently, they have become more easily available with the development of modern genetic engineering techniques. Here, we review existing mouse models for studying the Mediator, and data obtained in corresponding experiments. Full article
(This article belongs to the Special Issue Transgenic and Genetically Engineered Animal and Cell Culture Models)
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