Stable transgene expression in mice generated from retrovirally transduced embryonic stem cells

Area of Science:

• * Molecular Biology
• * Genetics
• * Developmental Biology

Background:

• * Gene silencing in retroviral vectors hinders transgenic animal production.
• * Stable gene expression is crucial for generating reliable transgenic models.

Purpose of the Study:

• * To develop a retroviral vector system for stable gene transfer and expression in mice.
• * To demonstrate the efficacy of the GCDsap vector for generating transgenic mice with EGFP expression.

Main Methods:

• * Utilized a murine stem cell virus-type retroviral vector (GCDsap) with modified LTR and PBS.
• * Employed vesicular stomatitis virus G protein pseudotyping to enhance viral titer.
• * Generated chimeric mice from retrovirally transduced embryonic stem cells for EGFP expression analysis.

Main Results:

• * Stable expression of the enhanced green fluorescent protein (EGFP) gene was achieved in chimeric mice.
• * EGFP expression was detected in diverse tissues, including hematopoietic cells, neurons, cardiac muscle, and intestine.
• * High EGFP expression persisted in progeny, indicating successful germline transmission of active proviruses, with 100% of leukocytes expressing EGFP.

Conclusions:

• * The described gene transfer system enables stable gene expression in transgenic animals.
• * This system is a valuable tool for generating transgenic mice and potentially manipulating human embryonic stem cells.