[Generation of bone morphogenetic protein 4 conditional RNA interference mice].
To explore bone morphogenetic protein 4 (BMP4) function in the developing bone, a BMP4 conditional RNA interference (CRNAi) vector was constructed based on the pBSK/U6 vector with a LoxPneo cassette. The transgene fragment targeting bmp4 was obtained by Kpn and Afl double digestion and was purified before being microinjected into fertilized eggs from FVB/NJ mice. BMP4CRNAi transgenic mice were genotyped by PCR. And the PCR positive mice were crossed with Col2a1-Cre transgenic mice, whose Cre recombinase was specifically expressed in osteo-chondro-progenitor cells. Bmp4 mRNA expression in primary chondrocytes were examined by semi-quantitive RT-PCR to determine RNA interference efficiency. Results showed that BMP4(CRNAi) mice and BMP4 (Col2a1-CRNAi) mice were produced successfully, and bmp4 knockdown efficiency in primary chondrocytes of BMP4 Col2a1-CRNAi mice was 81%. This transgenic mouse line provides excellent model for studying the role of BMP4 in chondrocyte development, and BMP4CRNAi mouse may be a good model for studying the role of BMP4 in different cells, tissues and organs through crossing with different Cre transgenic mice.
Li
Fu-Bing
FB
Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042, China.
DU
Xiao-Lan
XL
Yu
Ying
Y
Zhao
Ling
L
He
Qi-Fen
QF
Chen
Lin
L
D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D008957:Models, Genetic; D016133:Polymerase Chain Reaction; D034622:RNA Interference / Q000502:physiology*
D055415:Bone Morphogenetic Protein 4