NIEHS/NIH
2 ARTICLES PUBLISHED IN JoVE
Biology
Trans-inner Cell Mass Injection of Embryonic Stem Cells Leads to Higher Chimerism Rates
Gregory J. Scott 1, Artiom Gruzdev 1, Thomas B. Hagler 1, Manas K. Ray 1
1Knock Out Mouse Core, NIEHS/NIH
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 Goulding 1, Shih-Heng Chen 1, Mitzie Walker 1, Thomas M. Porter 1, Lucas Van Gorder 1, Amanda Mathew 1, Artiom Gruzdev 3, Erica Scappini 4, Charles Romeo 1
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.
