4.7 Article

Production of Genetically Modified Porcine Embryos via Lipofection of Zona-Pellucida-Intact Oocytes Using the CRISPR/Cas9 System

Related references

Note: Only part of the references are listed.
Article Veterinary Sciences

Zona pellucida treatment before CRISPR/Cas9-mediated genome editing of porcine zygotes

Zhao Namula et al.

Summary: The study showed that weakening the ZP does not affect the developmental competence, mutation rate, or mutation efficiency of electroporated zygotes, while ZP removal has a detrimental effect on embryonic development but may increase the mutation rate.

VETERINARY MEDICINE AND SCIENCE (2022)

Article Biochemistry & Molecular Biology

Effect of Aphidicolin, a Reversible Inhibitor of Eukaryotic Nuclear DNA Replication, on the Production of Genetically Modified Porcine Embryos by CRISPR/Cas9

Sergio Navarro-Serna et al.

Summary: This study evaluated the use of reversible inhibition of DNA replication to reduce mosaicism in embryos edited by CRISPR/Cas9. The findings showed a concentration-dependent DNA replication inhibition, leading to a decrease in mosaicism but a detrimental effect on embryo development. Balancing the use of this approach is necessary considering the impact on embryo development.

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022)

Article Medicine, Research & Experimental

Progressive genetic modifications of porcine cardiac xenografts extend survival to 9 months

Muhammad M. Mohiuddin et al.

Summary: This study reports the survival of genetically engineered porcine cardiac xenografts in baboon recipients for almost 9 months. The selective multi-gene modifications significantly improve the survival of cardiac xenografts, offering hope for transplantation in humans.

XENOTRANSPLANTATION (2022)

Article Chemistry, Multidisciplinary

Lipid-peptide nanocomplexes for mRNA delivery in vitro and in vivo

Dania Grant-Serroukh et al.

Summary: Despite recent advances in mRNA therapy, the lack of safe and effective delivery vehicles remains a major challenge. In this study, lipid-peptide nanocomplexes were systematically optimized for mRNA delivery. The optimized formulation showed improved transfection efficiency in vitro and in vivo, and retained stable physicochemical properties for a prolonged period of time. This lipid-peptide nanocomplex provides a promising platform for the delivery of therapeutic mRNA.

JOURNAL OF CONTROLLED RELEASE (2022)

Article Agriculture, Dairy & Animal Science

Gene editing in porcine embryos using a combination of electroporation and transfection methods

Koki Takebayashi et al.

Summary: CRISPR/Cas9 technology has greatly influenced the efficiency and effectiveness of genetic modifications. The study found that electroporation alone significantly increased the frequency of biallelic mutations, while lipofection alone required the removal of the zona pellucida to enable CRISPR/Cas9-based genome editing. Additionally, additional lipofectamine treatment did not improve the mutation rates.

REPRODUCTION IN DOMESTIC ANIMALS (2022)

Article Reproductive Biology

Generation of Calpain-3 knock-out porcine embryos by CRISPR-Cas9 electroporation and intracytoplasmic microinjection of oocytes before insemination

Sergio Navarro-Serna et al.

Summary: This study successfully generated a pig model of limb girdle muscular dystrophy type R1 (LGMDR1) by optimizing porcine oocyte electroporation. Electroporation, as a faster and easier method compared to microinjection, showed comparable rates of embryo development and mutation parameters.

THERIOGENOLOGY (2022)

Review Biochemistry & Molecular Biology

Advances in pig models of human diseases

Naipeng Hou et al.

Summary: Animal models play a critical role in medical research, and pigs are a preferred choice for modeling human diseases due to their similarity to humans in anatomy and physiology. Recent advances in genetic engineering have facilitated the rapid development of pig models for studying human diseases. This review provides an overview of the current status of pig models for human cardiovascular, metabolic, neurodegenerative, various genetic diseases, and xenotransplantation, and discusses areas for improvement.

ANIMAL MODELS AND EXPERIMENTAL MEDICINE (2022)

Article Agriculture, Dairy & Animal Science

Generation of CD163-edited pig via electroporation of the CRISPR/Cas9 system into porcine in vitro-fertilized zygotes

Fuminori Tanihara et al.

Summary: This study successfully generated CD163-modified pigs by introducing the CD163 gene into porcine zygotes using the CRISPR/Cas9 system. Three out of the four tested gRNAs showed high editing efficiency, but further improvements are needed for generating genetically modified pigs with high efficiency.

ANIMAL BIOTECHNOLOGY (2021)

Article Pharmacology & Pharmacy

Delivery of Protein Kinase A by CRISPRMAX and Its Effects on Breast Cancer Stem-Like Properties

Jun-Nian Zhou et al.

Summary: In this study, we successfully delivered PKA to breast cancer cells using the CRISPRMAX reagent, demonstrating its potential to inhibit EMT, affect mammosphere size, and impact chemoresistance. Direct enzyme delivery may be a promising strategy for inhibiting EMT/CSC-associated traits.

PHARMACEUTICS (2021)

Article Multidisciplinary Sciences

Protocol for assessment of the efficiency of CRISPR/Cas RNP delivery to different types of target cells

Marina A. Tyumentseva et al.

Summary: Efficiency and optimization of current CRISPR/Cas RNP delivery protocols to target cells will be assessed in this study using molecular cloning, protein expression and purification, cell culture, flow cytometry, and cellular imaging techniques. This will be the first extensive comparative study of popular CRISPR/Cas RNP delivery methods to human cell lines and primary cells.

PLOS ONE (2021)

Article Agriculture, Dairy & Animal Science

Lipofection-Mediated Introduction of CRISPR/Cas9 System into Porcine Oocytes and Embryos

Maki Hirata et al.

Summary: Liposome-mediated gene transfer is a feasible method for establishing a gene targeting framework and producing mutant animals even in laboratories without specialized equipment. However, the efficiency and factors affecting the system in mammalian oocytes and embryos remain unclear.

ANIMALS (2021)

Article Multidisciplinary Sciences

Generation of mutant pigs by lipofection-mediated genome editing in embryos

Maki Hirata et al.

Summary: The study developed a novel lipofection-mediated RNP transfection technique for generating gene-edited pigs without the need for specialized equipment, producing no detectable off-target events. Despite the need for improvements in editing efficiency, this practical method for rapid and large-scale gene editing in pigs has important agricultural and biomedical applications.

SCIENTIFIC REPORTS (2021)

Article Genetics & Heredity

Generation of Nonmosaic, Two-Pore Channel 2 Biallelic Knockout Figs in One veneration by CRISPR-Cas9 Microinjection Before Oocyte insemination

Sergio Navarro-Serna et al.

Summary: Studies have shown TPCs play roles in various pathophysiological processes, TPC2 KO pigs were generated using CRISPR-Cas9 for the first time to study TPC2's roles, and the timing of microinjection was found to greatly reduce mosaicism. Nonmosaic animals generated through this method facilitate germ line transmission of mutations for rapid and efficient generation of KO animal lines.

CRISPR JOURNAL (2021)

Article Biology

Timing and duration of lipofection-mediated CRISPR/Cas9 delivery into porcine zygotes affect gene-editing events

Qingyi Lin et al.

Summary: Lipofection-mediated introduction of the CRISPR/Cas9 system in porcine zygotes is a feasible gene editing method, but further improvements are needed to enhance editing efficiency. Timing and duration of lipofection play a critical role in the gene editing outcome, with longer incubation times leading to decreased mutation rates and blastocyst formation rates.

BMC RESEARCH NOTES (2021)

Article Multidisciplinary Sciences

Targeting of Uropathogenic Escherichia coli papG gene using CRISPR-dot nanocomplex reduced virulence of UPEC

Surbhi Gupta et al.

Summary: Urinary tract infections are common infectious diseases, but treating them is becoming difficult due to antibiotic resistance. This study developed a novel CRISPR gene editing strategy to target bacterial adhesion molecules, reducing pathogenicity.

SCIENTIFIC REPORTS (2021)

Article Cell Biology

Improved CRISPR/Cas9 gene editing in primary human myoblasts using low confluency cultures on Matrigel

Hayley Goullee et al.

Summary: This study demonstrates a simple and effective method to maximize CRISPR/Cas9-mediated gene editing in primary human myoblasts by delivering Cas9 ribonucleoprotein complexes on Matrigel-coated wells at low confluency. This approach achieved significantly higher editing efficiency compared to commercial recommendations, showing potential for improving genetic manipulation of skeletal muscle cells and other cell types.

SKELETAL MUSCLE (2021)

Article Agriculture, Dairy & Animal Science

CRISPR/Cas9-mediatedMSTNdisruption accelerates the growth of Chinese Bama pigs

Xiang-Xing Zhu et al.

REPRODUCTION IN DOMESTIC ANIMALS (2020)

Article Medicine, Research & Experimental

A porcine model of phenylketonuria generated by CRISPR/Cas9 genome editing

Erik A. Koppes et al.

JCI INSIGHT (2020)

Article Biochemistry & Molecular Biology

Deletion of CD163 Exon 7 Confers Resistance to Highly Pathogenic Porcine Reproductive and Respiratory Viruses on Pigs

Haitao Wang et al.

INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES (2019)

Article Multidisciplinary Sciences

Quadrupling efficiency in production of genetically modified pigs through improved oocyte maturation

Ye Yuan et al.

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA (2017)

Letter Biotechnology & Applied Microbiology

Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus

Kristin M. Whitworth et al.

NATURE BIOTECHNOLOGY (2016)

Article Biotechnology & Applied Microbiology

Improved delivery of Cas9 protein/gRNA complexes using lipofectamine CRISPRMAX

Xin Yu et al.

BIOTECHNOLOGY LETTERS (2016)

Article Biochemistry & Molecular Biology

Breaking-Cas-interactive design of guide RNAs for CRISPR-Cas experiments for ENSEMBL genomes

Juan C. Oliveros et al.

NUCLEIC ACIDS RESEARCH (2016)

Article Multidisciplinary Sciences

High-throughput genotyping of CRISPR/Cas9-mediated mutants using fluorescent PCR-capillary gel electrophoresis

Muhammad Khairul Ramlee et al.

SCIENTIFIC REPORTS (2015)

Article Veterinary Sciences

Characteristics and EGFP expression of porcine mammary gland epithelial cells

Yue-Mao Zheng et al.

RESEARCH IN VETERINARY SCIENCE (2010)

Article Agriculture, Dairy & Animal Science

Evaluation of RNA interference in developing porcine granulosa cells using fluorescence reporter genes

T Hirano et al.

JOURNAL OF REPRODUCTION AND DEVELOPMENT (2004)