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Enhanced transgenesis by intracytoplasmic injection of envelope‐free lentivirus
Author(s) -
Yang ShangHsun,
Agca Yuksel,
Cheng PeiHsun,
Yang JinJing,
Agca Cansu,
Chan Anthony Wing Sang
Publication year - 2007
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.20289
Subject(s) - transgenesis , biology , transgene , microbiology and biotechnology , lentivirus , viral envelope , green fluorescent protein , virology , embryo , virus , oocyte , gene , genetics , oogenesis , viral disease , glycoprotein
Abstract We demonstrate enhanced transgenesis in mice by intracytoplasmic injection of envelope‐free lentivirus. Envelope‐free lentivirus carrying the green fluorescent protein (GFP) gene under the control of the ubiquitin promoter (LVU‐GFP) was microinjected into the cytoplasm of mouse zygotes prior to embryo transfer. Ninety‐seven percent (31/32) of the adult mice were confirmed transgenic by PCR and Southern blot analysis; all founder mice express GFP when tail snips were examined by fluorescent microscopy prior to genomic DNA extraction. Transgene insertion numbers ranging from 1 to 32 were revealed by Southern blot analysis. Germline transmission was confirmed by the presence of transgene in F1 offspring. As expected, a lower transgenic rate (2.2%; 1/46) resulted when envelope‐free LVU‐GFP was microinjected into the perivitelline space (PVS) because cell recognition followed by membrane fusion between the viral envelope and the target cell is prerequisite for successful infection by envelope viruses. Here we demonstrate the competence of envelope‐free lentivirus in establishing stable gene integration by germline transgenesis in mice at high efficiency, by intracytoplasmic viral injection (INVI) of envelope‐free lentivirus into mouse zygotes. genesis 45:177–183, 2007. Published 2007 Wiley‐Liss, Inc.