[Generation of bone morphogenetic protein 4 conditional RNA inter-ference mice.]
March 16th, 2008 | by admin |[Generation of bone morphogenetic protein 4 conditional RNA inter-ference 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 BMP4CRNAi 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 FB, DU XL, Yu Y, Zhao L, He QF, Chen L.
State Key Laboratory of Trauma, Burns and Combined Injury, Trauma Center, Institute of Surgery Research, Daping Hospital, Third Mili-tary Medical University, Chongqing 400042, China E-mail: abbilson@yahoo.com.cn.