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 journalArticle

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 1

Fingerprint

Gene Transfer Techniques
Lentivirus
Green Fluorescent Proteins
Injections
Zygote
Southern Blotting
Transgenes
Virus Internalization
Embryo Transfer
Virus Diseases
Ubiquitin
Mental Competency
Genes
Tail
Microscopy
Cytoplasm
Polymerase Chain Reaction
DNA

All Science Journal Classification (ASJC) codes

  • Genetics
  • Endocrinology
  • Cell Biology

Cite this

Yang, S-H., Agca, Y., Cheng, P. H., Yang, J. J., Agca, C., & Chan, A. W. S. (2007). Enhanced transgenesis by intracytoplasmic injection of envelope-free lentivirus. Genesis, 45(4), 177-183. https://doi.org/10.1002/dvg.20289
Yang, Shang-Hsun ; Agca, Yuksel ; Cheng, Pei Hsun ; Yang, Jin Jing ; Agca, Cansu ; Chan, Anthony Wing Sang. / Enhanced transgenesis by intracytoplasmic injection of envelope-free lentivirus. In: Genesis. 2007 ; Vol. 45, No. 4. pp. 177-183.
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Yang, S-H, Agca, Y, Cheng, PH, Yang, JJ, Agca, C & Chan, AWS 2007, 'Enhanced transgenesis by intracytoplasmic injection of envelope-free lentivirus', Genesis, vol. 45, no. 4, pp. 177-183. https://doi.org/10.1002/dvg.20289

Enhanced transgenesis by intracytoplasmic injection of envelope-free lentivirus. / Yang, Shang-Hsun; Agca, Yuksel; Cheng, Pei Hsun; Yang, Jin Jing; Agca, Cansu; Chan, Anthony Wing Sang.

In: Genesis, Vol. 45, No. 4, 01.04.2007, p. 177-183.

Research output: Contribution to journalArticle

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