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2 ARTICLES PUBLISHED IN JoVE

Biology

Trans-inner Cell Mass Injection of Embryonic Stem Cells Leads to Higher Chimerism Rates

Gregory J. Scott¹, Artiom Gruzdev¹, Thomas B. Hagler¹, Manas K. Ray¹

Here we present a protocol to increase chimera production without the use of new equipment. A simple orientation change of the embryo for injection can increase the number of embryos produced, and potentially reduce the timeline to germline transmission.

JoVE Journal

Laser-assisted Lentiviral Gene Delivery to Mouse Fertilized Eggs

Negin P. Martin^*1, Page Myers^*2, Eugenia Goulding¹, Shih-Heng Chen¹, Mitzie Walker¹, Thomas M. Porter¹, Lucas Van Gorder¹, Amanda Mathew¹, Artiom Gruzdev³, Erica Scappini⁴, Charles Romeo¹

¹Neurobiology Laboratory, National Institute of Environmental Health Sciences, ²Comparative Medicine Branch, National Institute of Environmental Health Sciences, ³Reproductive and Developmental Biology Laboratory, National Institute of Environmental Health Sciences, ⁴Signal Transduction Laboratory, National Institute of Environmental Health Sciences

Mouse fertilized eggs and early stage embryos are protected by the zona pellucida, a glycoprotein matrix that forms a barrier against gene delivery. This article describes a protocol for perforating the zona with a laser to transduce embryonic cells with lentiviral vectors and to create transgenic mice.