Pseudotyped retroviruses for infecting axolotl.

Abstract

The ability to introduce DNA elements into host cells and analyze the effects has revolutionized modern biology. Here we describe a protocol to generate Moloney murine leukemia virus (MMLV)-based, replication-incompetent pseudotyped retrovirus capable of infecting axolotls and incorporating genetic information into their genome. When pseudotyped with vesicular stomatitis virus (VSV)-G glycoprotein, the retroviruses can infect a broad range of proliferative axolotl cell types. However, if the retrovirus is pseudotyped with an avian sarcoma leukosis virus (ASLV)-A envelope protein, only axolotl cells experimentally manipulated to express the cognate tumor virus A (TVA) receptor can be targeted by infections. These strategies enable robust transgene expression over many cell divisions, cell lineage tracing, and cell subtype targeting for gene expression.

• Authors: Tzu-Hsing Kuo, Jessica L Whited
• Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol) Vol. 1290 Pg. 127-40 ( 2015) ISSN: 1940-6029 [Electronic] United States
• PMID: 25740482 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Topics

• Alpharetrovirus (genetics)
• Ambystoma mexicanum (embryology, virology)
• Animals
• Extremities (embryology)
• Genetic Vectors (genetics)
• HEK293 Cells
• Humans
• Moloney murine leukemia virus (genetics, physiology)
• Open Reading Frames (genetics)
• Plasmids (genetics)
• Transfection (methods)
• Viral Load