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Targeting gene expression in the preimplantation mouse embryo using morpholino antisense oligonucleotides
Author(s) -
Siddall Laura S.,
Barcroft Lisa C.,
Watson Andrew J.
Publication year - 2002
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.10202
Subject(s) - biology , morpholino , embryo , oligonucleotide , gene expression , gene , genetics , microbiology and biotechnology , andrology , computational biology , gene knockdown , medicine
Morpholino antisense oligonucleotides act by blocking translation of their target gene products and are effective tools for down‐regulating gene expression. The current study was conducted to define treatment conditions for the use of morpholino oligonucleotides (MOs) in mammalian preimplantation embryos, and to employ MOs to target genes and study gene function in the early embryo. For the first time, ethoxylated polyethylenimine (EPEI), Lipofectin™ or Lysolecithin delivery agents were employed in combination with a fluorescent control MO and an α‐catenin specific MO, to down‐regulate gene expression during murine preimplantation development. Experiments applied to both two‐ and eight‐cell stage murine preimplantation embryos contrasted the efficacy of MO concentrations of 1, 2, 5, 10, and 20 µM and treatment delivery times of 3, 6, 24, and 48 hr. Continuous treatment of two‐cell embryos with Lipofectin and 20 µM α‐catenin MO for 48 hr resulted in a significant ( P < 0.05) reduction in development to the blastocyst stage and was accompanied by a marked reduction in α‐catenin protein. These results indicate that morpholino antisense oligonucleotides are effective tools for down‐regulating gene expression during mammalian preimplantation development. Mol. Reprod. Dev. 63: 413–421, 2002. © 2002 Wiley‐Liss, Inc.