Submit to Genes Review for Genes Propose a Special Issue

Journal Browser

Application of New Gene Editing Techniques in Pig Breeding

Print Special Issue Flyer
Special Issue Editors
Special Issue Information
Keywords
Published Papers

A special issue of Genes (ISSN 2073-4425). This special issue belongs to the section "Technologies and Resources for Genetics".

Deadline for manuscript submissions: closed (20 October 2022) | Viewed by 14137

Share This Special Issue

Special Issue Editor

Prof. Dr. Bo Zuo

E-Mail Website
Guest Editor

Department of Animal Science and Technology, Huazhong Agricultural University, Wuhan, China
Interests: gene editing; pig breed

Special Issue Information

Dear Colleagues,

We would like to invite you to participate in this Special Issue entitled Application of Gene Editing Techniques in Pig Breeding.

Pigs are among the most important livestock for meat production. With the development of gene editing techniques and the identification of major genes affecting economically important traits, genetic engineering in livestock has been widely applied for disease resistance and genetic improvement of production traits. Compared with other gene editing techniques, CRISPR/Cas9 systems are powerful tools for gene editing and allow for the creation of new genotypes by changing the target loci that introduce the locus double strand breaks and addressed by endogenous repair pathways. To date, gene edited pigs such as MSTN and CD163 gene edited pigs have been used to breed new strains of pigs with disease resistance and higher productivity in agriculture. Besides, pigs are regarded as ideal biomedical animals owing to their similarity in anatomy, physiology, and organ size to humans, and many pig models of human diseases have been established for drug screening and pathogenic mechanisms. In this Special Issue, we invite researchers to submit articles, reviews, and prospects about gene editing techniques and their applications in pig breeding and biomedical research.

Prof. Dr. Bo Zuo
Guest Editor

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. Genes is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). 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

pig
gene editing
CRISPR/Cas9
economically important traits, disease resistance
major genes
breeding
biomedical research
genetic improvement

Published Papers (5 papers)

Download All Papers

Research

Jump to: Review

20 pages, 3348 KiB

Open AccessArticle

Simulating the Commercial Implementation of Gene-Editing for Influenza A Virus Resistance in Pigs: An Economic and Genetic Analysis

by Hamish A. Salvesen, Timothy J. Byrne, C. Bruce A. Whitelaw and Fiona S. Hely

Genes 2022, 13(8), 1436; https://0-doi-org.brum.beds.ac.uk/10.3390/genes13081436 - 12 Aug 2022

Cited by 2 | Viewed by 2214

Abstract

The development of swine Influenza A Virus resistance along with genetic technologies could complement current control measures to help to improve animal welfare standards and the economic efficiency of pig production. We have created a simulation model to assess the genetic and economic implications of various gene-editing methods that could be implemented in a commercial, multi-tiered swine breeding system. Our results demonstrate the length of the gene-editing program was negatively associated with genetic progress in commercial pigs and that the time required to reach fixation of resistance alleles was reduced if the efficiency of gene-editing is greater. The simulations included the resistance conferred in a digenic model, the inclusion of genetic mosaicism in progeny, and the effects of selection accuracy. In all scenarios, the level of mosaicism had a greater effect on the time required to reach resistance allele fixation and the genetic progress of the herd than gene-editing efficiency and zygote survival. The economic analysis highlights that selection accuracy will not affect the duration of gene-editing and the investment required compared to the effects of gene-editing-associated mosaicism and the swine Influenza A Virus control strategy on farms. These modelling results provide novel insights into the economic and genetic implications of targeting two genes in a commercial pig gene-editing program and the effects of selection accuracy and mosaicism. Full article

(This article belongs to the Special Issue Application of New Gene Editing Techniques in Pig Breeding)

► Show Figures

Figure 1

10 pages, 5309 KiB

Open AccessArticle

Identification of the CKM Gene as a Potential Muscle-Specific Safe Harbor Locus in Pig Genome

by Youcai Xiong, Rongzhi Zhuang, Guangxing Zhao, Yanwen Liu, Yinyu Su, Wei Wang, Xiaoning Xi, Yanyu Yang, Xiaosong Han, Shengsong Xie, Heng Wang, Xinyun Li, Bo Zuo, Shuhong Zhao, Zheng Feng and Jinxue Ruan

Genes 2022, 13(5), 921; https://0-doi-org.brum.beds.ac.uk/10.3390/genes13050921 - 21 May 2022

Cited by 2 | Viewed by 2093

Abstract

Genetically modified pigs have shown considerable application potential in the fields of life science research and livestock breeding. Nevertheless, a barrier impedes the production of genetically modified pigs. There are too few safe harbor loci for the insertion of foreign genes into the pig genome. Only a few loci (pRosa26, pH11 and Pifs501) have been successfully identified to achieve the ectopic expression of foreign genes and produce gene-edited pigs. Here, we use CRISPR/Cas9-mediated homologous directed repair (HDR) to accurately knock the exogenous gene-of-interest fragments into an endogenous CKM gene in the porcine satellite cells. After porcine satellite cells are induced to differentiate, the CKM gene promoter simultaneously initiates the expression of the CKM gene and the exogenous gene. We infer preliminarily that the CKM gene can be identified as a potential muscle-specific safe harbor locus in pigs for the integration of exogenous gene-of-interest fragments. Full article

(This article belongs to the Special Issue Application of New Gene Editing Techniques in Pig Breeding)

► Show Figures

Figure 1

13 pages, 2491 KiB

Open AccessArticle

An Inexpensive CRISPR-Based Point-of-Care Test for the Identification of Meat Species and Meat Products

by Dagang Tao, Xiao Xiao, Xiaochen Lan, Bingrong Xu, Yuan Wang, Emmanuel Mulaya Khazalwa, Wenya Pan, Jinxue Ruan, Yu Jiang, Xiangdong Liu, Changchun Li, Ruizhen Ye, Xinyun Li, Jing Xu, Shuhong Zhao and Shengsong Xie

Genes 2022, 13(5), 912; https://0-doi-org.brum.beds.ac.uk/10.3390/genes13050912 - 19 May 2022

Cited by 8 | Viewed by 2416

Abstract

The growing demand for and supply of meat and meat products has led to a proportional increase in cases of meat adulteration. Adulterated meat poses serious economic and health consequences globally. Current laboratory methods for meat species identification require specialized equipment with limited field applications. This study developed an inexpensive, point-of-care Loop-Mediated Isothermal Amplification (LAMP)-CRISPR/Cas12a colorimetric assay to detect meat species using a Texas Red-labelled single-strand (ssDNA) reporter. As low as 1.0 pg/µL of the porcine NADH4, the chicken NADH dehydrogenase subunit 2 (ND2) and the duck D-loop genes was detectable under white, blue and ultraviolet light. The test turnaround time from DNA extraction to visualization was approximately 40 min. The assay accurately detected pure and mixed-meat products in the laboratory (n = 15) and during a pilot point-of-care test (n = 8) in a food processing factory. The results are 100% reproducible using lateral flow detection strips and the real-time PCR detection instrument. This technology is fully deployable and usable in any standard room. Thus, our study demonstrates that this method is a straightforward, specific, sensitive, point-of-care test (POCT) adaptable to various outlets such as customs, quarantine units and meat import/export departments. Full article

(This article belongs to the Special Issue Application of New Gene Editing Techniques in Pig Breeding)

► Show Figures

Figure 1

15 pages, 2576 KiB

Open AccessArticle

Efficient Simultaneous Introduction of Premature Stop Codons in Three Tumor Suppressor Genes in PFFs via a Cytosine Base Editor

by Haoyun Jiang, Qiqi Jing, Qiang Yang, Chuanmin Qiao, Yaya Liao, Weiwei Liu and Yuyun Xing

Genes 2022, 13(5), 835; https://0-doi-org.brum.beds.ac.uk/10.3390/genes13050835 - 07 May 2022

Cited by 2 | Viewed by 2043

Abstract

Base editing is an efficient and precise gene-editing technique, by which a single base can be changed without introducing double-strand breaks, and it is currently widely used in studies of various species. In this study, we used hA3A-BE3-Y130F to simultaneously introduce premature stop codons (TAG, TGA, and TAA) into three tumor suppressor genes, TP53, PTEN, and APC, in large white porcine fetal fibroblasts (PFFs). Among the isolated 290 single-cell colonies, 232 (80%) had premature stop codons in all the three genes. C−to−T conversion was found in 98.6%, 92.8%, and 87.2% of these cell colonies for TP53, PTEN, and APC, respectively. High frequencies of bystander C−to−T edits were observed within the editing window (positions 3–8), and there were nine (3.01%) clones with the designed simultaneous three-gene C−to−T conversion without bystander conversion. C−to−T conversion outside the editing window was found in 9.0%, 14.1%, and 26.2% of the 290 cell colonies for TP53, PTEN, and APC, respectively. Low-frequency C−to−G or C−to−A transversion occurred in APC. The mRNA levels of the three genes showed significant declines in triple-gene-mutant (Tri-Mut) cells as expected. No PTEN and a significantly lower (p < 0.05) APC protein expression were detected in Tri-Mut cells. Interestingly, the premature stop codon introduced into the TP53 gene did not eliminate the expression of its full-length protein in the Tri-Mut cells, suggesting that stop codon read-through occurred. Tri-Mut cells showed a significantly higher (p < 0.05) proliferation rate than WT cells. Furthermore, we identified 1418 differentially expressed genes (DEGs) between the Tri-Mut and WT groups, which were mainly involved in functions such as tumor progression, cell cycle, and DNA repair. This study indicates that hA3A-BE3-Y130F can be a powerful tool to create diverse knockout cell models without double-strand breaks (DSBs), with further possibilities to produce porcine models with various purposes. Full article

(This article belongs to the Special Issue Application of New Gene Editing Techniques in Pig Breeding)

► Show Figures

Figure 1

Review

Jump to: Research

18 pages, 1290 KiB

Open AccessReview

Double-Stranded Break Repair in Mammalian Cells and Precise Genome Editing

by Akhtar Ali, Wei Xiao, Masroor Ellahi Babar and Yanzhen Bi

Genes 2022, 13(5), 737; https://0-doi-org.brum.beds.ac.uk/10.3390/genes13050737 - 22 Apr 2022

Cited by 5 | Viewed by 4610

Abstract

In mammalian cells, double-strand breaks (DSBs) are repaired predominantly by error-prone non-homologous end joining (NHEJ), but less prevalently by error-free template-dependent homologous recombination (HR). DSB repair pathway selection is the bedrock for genome editing. NHEJ results in random mutations when repairing DSB, while HR induces high-fidelity sequence-specific variations, but with an undesirable low efficiency. In this review, we first discuss the latest insights into the action mode of NHEJ and HR in a panoramic view. We then propose the future direction of genome editing by virtue of these advancements. We suggest that by switching NHEJ to HR, full fidelity genome editing and robust gene knock-in could be enabled. We also envision that RNA molecules could be repurposed by RNA-templated DSB repair to mediate precise genetic editing. Full article

(This article belongs to the Special Issue Application of New Gene Editing Techniques in Pig Breeding)

► Show Figures