title: Small interference RNA-mediated knockdown of sperm associated antigen 9 having structural homology with c-Jun N-terminal kinase-interacting protein.
abstract: Recently, we reported a novel testis-specific sperm associated antigen 9 (SPAG9) protein, a new member of the JNK-interacting protein family, having a functional role in sperm-egg fusion [N. Jagadish, R. Rana, R. Selvi, D. Mishra, M. Garg, S. Yadav, J.C. Herr, K. Okumura, A. Hasegawa, K. Koyama, A. Suri, Biochem. J. 389 (2005) 73-82]. NCBI Blast searches revealed SPAG9 nucleotide sequence similarities with ESTs of various cancerous tissues. In the present study, we compared the efficiency of two independent SPAG9 specific small interfering RNA (siRNA) constructs, BS/U6/spag9 and BS/U6/spag9-I, to ablate the SPAG9 expression in mammalian cells. A positive correlation between the ratio of target gene versus siRNA and the suppression of SPAG9 expression was observed. Further, the cotransfection of BS/U6/spag9 with pcDNA-SPAG9 and pFlag-CMV2-JNK-3 resulted in specific suppression of SPAG9 without affecting JNK-3 expression. The present investigation will eventually extend the application of SPAG9 siRNA in in vivo targeting experiments that aim to define the SPAG9 functional genomics in tumor and reproductive biology.
authors.affiliation: Genes and Proteins Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, J.N.U. Campus, New Delhi-110 067, India.
authors.forename: Ritu
authors.initials: R
authors.lastname: Rana
authors.forename: Nirmala
authors.initials: N
authors.lastname: Jagadish
authors.forename: Manoj
authors.initials: M
authors.lastname: Garg
authors.forename: Deepshikha
authors.initials: D
authors.lastname: Mishra
authors.forename: Neetu
authors.initials: N
authors.lastname: Dahiya
authors.forename: Dipak
authors.initials: D
authors.lastname: Chaurasiya
authors.forename: Anil
authors.initials: A
authors.lastname: Suri
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D020868:Gene Silencing; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D034741:RNA, Small Interfering / Q000235:genetics*; D017386:Sequence Homology, Amino Acid
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D008565:Membrane Proteins; D034741:RNA, Small Interfering; C111047:SPAG9 protein, human; D048031:JNK Mitogen-Activated Protein Kinases