Pseudotyped retroviruses for infecting axolotl

Area of Science:

• Developmental Biology
• Genetics
• Molecular Biology

Background:

• Introducing genetic material into host cells is crucial for biological research.
• Axolotls are important model organisms for regeneration studies.

Purpose of the Study:

• To develop a method for introducing genetic elements into axolotl cells using retroviruses.
• To enable genetic manipulation and gene expression analysis in axolotls.

Main Methods:

• Generation of Moloney murine leukemia virus (MMLV)-based, replication-incompetent retroviruses.
• Pseudotyping retroviruses with vesicular stomatitis virus (VSV)-G glycoprotein for broad infection.
• Pseudotyping retroviruses with avian sarcoma leukosis virus (ASLV)-A envelope protein for receptor-specific infection (TVA receptor).

Main Results:

• Retroviruses pseudotyped with VSV-G efficiently infected various proliferative axolotl cell types.
• Retroviruses pseudotyped with ASLV-A selectively infected axolotl cells engineered to express the TVA receptor.
• Achieved robust transgene expression over multiple cell divisions.

Conclusions:

• Developed a versatile retroviral gene delivery system for axolotls.
• Enabled broad and targeted gene delivery strategies for axolotl cell manipulation.
• Facilitated cell lineage tracing and cell subtype-specific gene expression studies in axolotls.