Enhanced transgenesis by intracytoplasmic injection of envelope-free lentivirus

Shang Hsun Yang, Yuksel Agca, Pei Hsun Cheng, Jin Jing Yang, Cansu Agca, Anthony Wing Sang Chan

Research output: Contribution to journalArticlepeer-review

11 Citations (Scopus)

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 lentivifus 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.

Original languageEnglish
Pages (from-to)177-183
Number of pages7
JournalGenesis
Volume45
Issue number4
DOIs
Publication statusPublished - 2007 Apr

All Science Journal Classification (ASJC) codes

  • Genetics
  • Endocrinology
  • Cell Biology

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