Synergistic power of genomic selection, assisted reproductive technologies, and gene editing to drive genetic improvement of cattle

CABI Agriculture and Bioscience, Год журнала: 2022, Номер 3(1)

Опубликована: Март 4, 2022

Abstract Genetic improvement of cattle around the globe has been, and will continue to be, an important driver animal agriculture sustainability. There are several reproductive molecular biotechnologies that used in genetic cattle, their impact on rate progress is maximized when combined synergistically a structured breeding program with clear objective. One most recently developed increasingly popular tools, gene editing, allows breeders precisely add, delete, or replace letters code so as influence specific trait interest (e.g., disease resistance), little one generation. However, for editing be factor improvement, it must integrate smoothly into conventional programs maintain accelerate rates gain. This review first summarizes current state key available then discusses potential strategies effectively incorporating methods disseminating traits improved via editing. Moreover, examines how strategies, including use differ depending industry sector (i.e., dairy beef), region world which they being deployed.

Язык: Английский

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CRISPR/Cas9-editing of PRNP in Alpine goats

Veterinary Research, Год журнала: 2025, Номер 56(1)

Опубликована: Янв. 13, 2025

Abstract Misfolding of the cellular PrP (PrP c ) protein causes prion disease, leading to neurodegenerative disorders in numerous mammalian species, including goats. A lack induces complete resistance disease. The aim this work was engineer Alpine goats carrying knockout (KO) alleles PRNP , -encoding gene, using CRISPR/Cas9-ribonucleoproteins and single-stranded donor oligonucleotides. targeted region preceded Ter mutation previously described Norwegian Genome editors were injected under zona pellucida prior electroporation 565 goat embryos/oocytes. total 122 two-cell-stage embryos transferred 46 hormonally synchronized recipient Six remained pregnant naturally gave birth 10 offspring. Among newborns, eight founder animals genome-edited obtained. Eight different mutated observed, five inducing KO mutations. Three founders carried only phenotypically indistinguishable from their wild-type counterparts. them, one male a base pair insertion allele is currently used rapidly extend -KO line for future characterization. In addition alleles, del6 genetic variant has been identified one-third animals. This new will be tested its potential properties with respect Future studies also evaluate effects background on other characters associated KO, as breed or species.

Язык: Английский

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Retrospect and prospect: reproductive technologies in beef cattle

Molecular Biology Reports, Год журнала: 2025, Номер 52(1)

Опубликована: Фев. 20, 2025

Язык: Английский

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Multidrug-Resistant Microbial Therapy Using Antimicrobial Peptides and the CRISPR/Cas9 System

Veterinary Medicine Research and Reports, Год журнала: 2022, Номер Volume 13, С. 173 - 190

Опубликована: Авг. 1, 2022

Abstract: The emergence and spread of multidrug-resistant microbes become a serious threat to animal human health globally because their less responsiveness conventional antimicrobial therapy. Multidrug-resistant microbial infection poses higher morbidity mortality rate with significant economic losses. Currently, peptides the CRISPR/Cas9 system are explored as alternative therapy circumvent challenges organisms. Antimicrobial small molecular weight, cationic extracted from all living It is promising drug candidate for treatment by direct killing or indirectly modulating innate immune system. another novel used manage infection. versatile gene-editing tool that uses engineered single guide RNA targeted gene recognition Cas9 enzyme destruction target nucleic acids. Both were successfully treat nosocomial infections caused ESKAPE pathogens, which developed resistance various antimicrobials. Despite, valuable roles in treatments, both CRISPR/Cas systems have limitations like toxicity, instability, incurring high manufacturing costs. Thus, this review paper gives detailed explanations circumventing infections, its limitation prospects clinical applications. Keywords: peptides, applications, CRISPR/cas system, organism

Язык: Английский

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Global status of gene edited animals for agricultural applications

The Veterinary Journal, Год журнала: 2024, Номер 305, С. 106142 - 106142

Опубликована: Май 22, 2024

Gene editing (GnEd) involves using a site-directed nuclease to introduce double-strand break (DSB) at targeted location in the genome. A literature search was performed on use of GnEd animals for agricultural applications. Data extracted from 212 peer-reviewed articles that described production least one living animal employing technologies purposes. The most common system reported CRISPR/Cas9, and frequent type edit unguided insertion or deletion resulting repair DSB leading knock-out (KO) mutation. Animal groups included reviewed papers were ruminants (cattle, sheep, goats, n=63); monogastrics (pigs rabbit, n=60); avian (chicken, duck, quail, n=17); aquatic (many species, n=65), insects (honeybee, silkworm, n=7). Yield (32%), followed by reproduction (21%) disease resistance (17%) most- commonly traits. Over half had Chinese first-authorship. Several countries, including Argentina, Australia, Brazil, Colombia Japan, have adopted regulatory policy considers KO mutations introduced following as akin natural genetic variation, therefore treat these analogously those produced conventional breeding. This approach has resulted non-GMO determination small number food applications, three species fast-growing fish, (red sea bream, olive flounder tiger pufferfish Japan), fish cattle Argentina porcine reproductive respiratory syndrome (PRRS) virus disease-resistant pigs Colombia.

Язык: Английский

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Potentials, prospects and applications of genome editing technologies in livestock production

Saudi Journal of Biological Sciences, Год журнала: 2021, Номер 29(4), С. 1928 - 1935

Опубликована: Ноя. 24, 2021

In recent years, significant progress has been achieved in genome editing applications using new programmable DNA nucleases such as zinc finger (ZFNs), transcription activator-like endonucleases (TALENs) and the clustered regularly interspaced short palindromic repeats/Cas9 system (CRISPR/Cas9). These tools are capable of nicking precisely by targeting specific sequences, enable addition, removal or substitution nucleotides via double-stranded breakage at genomic loci. CRISPR/Cas system, one most tools, affords ability to efficiently generate multiple nicks single experiment. Moreover, systems relatively easy cost effective when compared other technologies. This is part because rely on RNA-DNA binding, unlike that protein-DNA interactions, which higher flexibility more fidelity. Genome have significantly contributed different aspects livestock production disease resistance, improved performance, alterations milk composition, animal welfare biomedicine. However, despite these contributions future potential, technologies also inherent risks, therefore, ethics social acceptance crucial factors associated with implementation review emphasizes impact development breeding numerous species cattle, pigs, sheep goats. discusses mechanisms behind technologies, their potential applications, risks should be considered context livestock.

Язык: Английский

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Optimizing oocyte electroporation for genetic modification of porcine embryos: Evaluation of the parthenogenetic activation

Theriogenology, Год журнала: 2024, Номер 218, С. 126 - 136

Опубликована: Фев. 1, 2024

In reproductive biology, understanding the effects of novel techniques on early embryo development is paramount importance. To date, electrical activation oocytes prior to in vitro fertilization (IVF) are not well understood. The aim this study was investigate oocyte electroporation IVF and differentiate between true embryos parthenotes by using a TPCN2 knock-out (KO) male evaluate presence KO allele resulting blastocysts. consisted three experiments. first one examined with without subsequent found that electroporated had higher rates, increased occurrence single pronucleus, no effect sperm penetration. Cleavage rates improved oocytes, but blastocyst remained constant. Genotype analysis revealed significant increase proportion groups compared control (30.2 % vs. 6.8 %). second experiment two media, Opti-MEM Nuclease-Free Duplex Buffer (DB). DB induced degeneration lower cleavage than Opti-MEM, while parthenogenetic formation consistent (60.0 48.5 third experiment, timing relative evaluated (1 h before IVF, immediately 7 after IVF). Electroporation resulted different pronuclear proportions other groups. penetration rate immediate group, all control. Blastocyst Genotyping differences parthenote among groups, these were (56.25 %, 63.89 51.61 2.44 respectively), showed mutation when performed IVF. Overall, comprehensive sheds light potential for creating genetically modified importance media selection process, best being more efficient regarding rate, post-IVF, even did differ Further studies needed reduce maintaining high optimize use procedure generation gene-edited pig oocyte/zygote electroporation.

Язык: Английский

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Electroporation Delivery of Cas9 sgRNA Ribonucleoprotein-Mediated Genome Editing in Sheep IVF Zygotes

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(17), С. 9145 - 9145

Опубликована: Авг. 23, 2024

The utilization of electroporation for delivering CRISPR/Cas9 system components has enabled efficient gene editing in mammalian zygotes, facilitating the development genome-edited animals. In this study, our research focused on targeting

Язык: Английский

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Efficient Generation of SOCS2 Knock-Out Sheep by Electroporation of CRISPR-Cas9 Ribonucleoprotein Complex with Dual-sgRNAs

The CRISPR Journal, Год журнала: 2025, Номер unknown

Опубликована: Янв. 14, 2025

In mice, naturally occurring and induced mutations in the suppressor of cytokine signaling-2 (Socs2) gene are associated with a high growth phenotype characterized by rapid post-weaning weight gain 30-50% heavier mature body weight. this work, we demonstrate an electroporation-based method producing SOCS2 knock-out (KO) sheep. Electroporation dual-guide CRISPR-Cas9 ribonucleoprotein complexes targeting was performed 6 h post-fertilization sheep zygotes. Fifty-two blastocysts were transferred to 13 estrus-synchronized recipients, yielding five live lambs one stillborn. These all carried predicted result KO. Three large deletion alleles which evaded detection initial PCR screening. Off-target analysis using whole genome sequencing comparing frequency regions within 100 bp possible sgRNA binding sites (up 4 mismatches) elsewhere showed no significant difference when unedited control edited animals (p = 0.71). conclusion, electroporation zygotes RNPs effective at generating substantial off-target activity.

Язык: Английский

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Optimizing Genome Editing in Bovine Cells: A Comparative Study of Cas9 Variants and CRISPR Delivery Methods

Biocatalysis and Agricultural Biotechnology, Год журнала: 2025, Номер unknown, С. 103553 - 103553

Опубликована: Март 1, 2025

Язык: Английский

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