In vivo genetic manipulation of the rat trophoblast cell lineage using lentiviral vector delivery

In this report, we have adapted a lentiviral gene delivery technique for genetic modification of the rat trophoblast cell lineage. Blastocysts were incubated with lentiviral particles and transferred into the uteri of pseudopregnant female rats, harvested at various times during gestation, and then...

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Bibliographic Details
Published in:Genesis (New York, N.Y. : 2000) N.Y. : 2000), 2009-07, Vol.47 (7), p.433-439
Main Authors: Lee, Dong-Soo, Rumi, M.A. Karim, Konno, Toshihiro, Soares, Michael J.
Format: Article
Language:English
Subjects:
Animals
Blotting, Western
Cell Lineage
Female
Genetic Vectors
green fluorescent protein
Green Fluorescent Proteins - genetics
lentivirus
Lentivirus - genetics
placenta
Pseudopregnancy
rat
Rats
Rats, Sprague-Dawley
RNA - genetics
trophoblast
Trophoblasts - cytology
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Summary:In this report, we have adapted a lentiviral gene delivery technique for genetic modification of the rat trophoblast cell lineage. Blastocysts were incubated with lentiviral particles and transferred into the uteri of pseudopregnant female rats, harvested at various times during gestation, and then analyzed. Two test systems were evaluated: (1) delivery of an enhanced green fluorescent protein (EGFP) gene under the control of constitutive promoters to rat blastocysts; (2) delivery of EGFP short hairpin RNA (shRNA) to rat blastocysts constitutively expressing EGFP. Lentiviral packaged gene constructs were efficiently and specifically delivered to all trophoblast cell lineages. Additionally, lentiviral mediated transfer of shRNAs was an effective strategy for modifying gene expression in trophoblast cell lineages. This technique will permit the in vivo evaluation of “gain‐of‐function” and “loss‐of‐function” manipulations in the rat trophoblast cell lineage. genesis 47:433–439, 2009. © 2009 Wiley‐Liss, Inc.
ISSN:1526-954X
1526-968X
DOI:10.1002/dvg.20518