Generating non-human primate models of human diseases is important for the development of therapeutic strategies especially for neurodegenerative diseases. The common marmoset has attracted attention as a new experimental animal model, and many transgenic marmosets have been produced using lentiviral vector-mediated transgenesis. However, lentiviral vectors have a size limitation of up to 8 kb in length for transgene applications. Therefore, the present study aimed to optimize a piggyBac transposon-mediated gene transfer method in which transgenes longer than 8 kb were injected into the perivitelline space of marmoset embryos, followed by electroporation. We constructed a long piggyBac vector carrying the gene responsible for Alzheimer's disease. The optimal weight ratio of the piggyBac transgene vector to the piggyBac transposase mRNA was examined using mouse embryos. Transgene integration into the genome was confirmed in 70.7% of embryonic stem cells established from embryos injected with 1000 ng of transgene and transposase mRNA. Under these conditions, long transgenes were introduced into marmoset embryos. All embryos survived after transgene introduction treatment, and transgenes were detected in 70% of marmoset embryos. The transposon-mediated gene transfer method developed in this study can be applied to the genetic modification of non-human primates, as well as large animals.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10256193PMC
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0287065PLOS

Publication Analysis

Top Keywords

marmoset embryos
16
transposon-mediated gene
12
gene transfer
12
transfer method
12
piggybac transposon-mediated
8
common marmoset
8
transposase mrna
8
embryos
7
marmoset
5
transgene
5

Similar Publications

Common and uncommon findings in marmoset and human trophoblast stem cells.

Cell Stem Cell

October 2024

Department of Informative Genetics, Tohoku University Graduate School of Medicine, Sendai 980-8575, Japan. Electronic address:

Article Synopsis
  • - Siriwardena et al. investigate the development of marmoset trophoblasts before and after implantation, focusing on early stages of embryo development.
  • - They successfully create marmoset trophoblast stem cell (TSC) lines from pluripotent stem cells (PSCs) for further study.
  • - A comparative analysis between marmoset and human TSCs reveals differences in implantation and placentation strategies unique to each species.
View Article and Find Full Text PDF

Marmoset and human trophoblast stem cells differ in signaling requirements and recapitulate divergent modes of trophoblast invasion.

Cell Stem Cell

October 2024

Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK; Centre for Trophoblast Research, University of Cambridge, Cambridge, UK; Wellcome Trust, Medical Research Council Stem Cell Institute, University of Cambridge, Cambridge, UK. Electronic address:

Article Synopsis
  • - The study investigates early human trophoblast development using marmoset embryos, bridging gaps in understanding due to the inaccessibility of human early conceptus.
  • - Researchers successfully created trophoblast stem cells (TSCs) from marmoset pluripotent stem cells, demonstrating unique characteristics and differentiation potential compared to human TSCs.
  • - The findings suggest that specific culture conditions for marmosets can maintain a trophoblast-like identity, revealing insights into evolutionary differences in implantation and enhancing knowledge of human development.
View Article and Find Full Text PDF

Background: The common marmoset, Callithrix jacchus, is an invaluable model in biomedical research. Its use includes genetic engineering applications, which require manipulations of oocytes and production of embryos in vitro. To maximize the recovery of oocytes suitable for embryo production and to fulfil the requirements of the 3R principles to the highest degree possible, optimization of ovarian stimulation protocols is crucial.

View Article and Find Full Text PDF

The sperm epigenome is thought to affect the developmental programming of the resulting embryo, influencing health and disease in later life. Age-related methylation changes in the sperm of old fathers may mediate the increased risks for reproductive and offspring medical problems. The impact of paternal age on sperm methylation has been extensively studied in humans and, to a lesser extent, in rodents and cattle.

View Article and Find Full Text PDF

Purpose: The common marmoset (Callithrix jacchus) provides an ideal model to study early development of primates, and an in vivo platform to validate conclusions from in vitro studies of human embryos and embryo models. Currently, however, no established staging atlas of marmoset embryonic development exists. Using high-resolution, longitudinal ultrasound scans on live pregnant marmosets, we present the first dynamic in vivo imaging of entire primate gestation beginning with attachment until the last day before birth.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!