Your browser doesn't support javascript.
loading
Construction of epithelia membrane protein 1 gene-deficient in human fetal nucleus pulposus cell model by lentivirus -mediated RNA interference / 中国骨伤
China Journal of Orthopaedics and Traumatology ; (12): 842-845, 2012.
Artículo en Chino | WPRIM | ID: wpr-313813
ABSTRACT
<p><b>OBJECTIVE</b>To construct Epithelia Membrane Protein 1 gene-deficient in human fetal nucleus pulposus model by lentivirus-mediated RNA interference for building a platform for illustrating the biomechanisms role of EMP-1 during human intervertebral disc degeneration.</p><p><b>METHODS</b>The lentivirus vector with shRNA targeting EMP-1 mRNA was transected into 293FT cells by liposome. Then the lentivirus supernatant was obtained and used for infecting human fetal nucleus pulposus. The expression of GFP was observed under fluorescence microscope after 48 h. The viral particles were collected at 72 h after transfection. The efficacy of gene interference was tested by Western blot and Real-time RT-PCR. Analysis the results of the fluorescent microscope scenes and get the average values of EMP-1/GAPDH by detected the interference efficiency of various interference DNA sequences with western blot and semi quantitative RT-PCR methods.</p><p><b>RESULTS</b>The lentivirns with high titer were obtained and the EMP-1 gene deficient cell strains were obtained. Semi quantitative RT-PCR and Western blot proved the average values of EMP-1/GAPDH decreased from 0.46 to 0.32 and 0.5 to 0.25 (P < 0.01).</p><p><b>CONCLUSION</b>Lentivirus packaging technology can be mastered skillfully. EMP-1 gene-deficient cell models are successfully established.</p>
Asunto(s)
Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Transfección / Lentivirus / Receptores de Superficie Celular / Interferencia de ARN / Células HEK293 / Feto / Genética / Disco Intervertebral / Metabolismo / Proteínas de Neoplasias Límite: Humanos Idioma: Chino Revista: China Journal of Orthopaedics and Traumatology Año: 2012 Tipo del documento: Artículo

Similares

MEDLINE

...
LILACS

LIS

Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Transfección / Lentivirus / Receptores de Superficie Celular / Interferencia de ARN / Células HEK293 / Feto / Genética / Disco Intervertebral / Metabolismo / Proteínas de Neoplasias Límite: Humanos Idioma: Chino Revista: China Journal of Orthopaedics and Traumatology Año: 2012 Tipo del documento: Artículo