Pseudotyped retroviruses for infecting axolotl.
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.
Brigham Regenerative Medicine Center, Department of Orthopedic Surgery, Brigham & Women's Hospital, Cambridge, MA, USA.
Tzu-Hsing
TH
Kuo
Jessica L
JL
Whited
D001355:Alpharetrovirus / Q000235:genetics; D000558:Ambystoma mexicanum / Q000196:embryology / Q000821:virology*; D000818:Animals; D005121:Extremities / Q000196:embryology; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008979:Moloney murine leukemia virus / Q000235:genetics* / Q000502:physiology*; D016366:Open Reading Frames / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D014162:Transfection / Q000379:methods*; D019562:Viral Load