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In Vitro knock out of Inducible Cyclicamp Early Repressor(ICER) in zebrafish cell lines through the use of the CrispR/Cas9 system (LB530)
Author(s) -
Hasani Aliaskar,
Molina Carlos
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.lb530
Subject(s) - biology , zebrafish , gene , crispr , repressor , microbiology and biotechnology , transcriptional regulation , genetics , transcription factor
ICER is a dominant negative transcriptional regulator induced by cAMP and it binds to CREB or CREM in order to repress cAMP‐mediated gene expression. Inhibin alpha, Inha, has been shown to regulate gonadal stromal cell proliferation and suppress tumor activity. The multi‐functions of ICER: as a neuronal plasticity, tumor suppressor, regulator of apoptosis, and etc., have led to the characterization of the relevant gene in various organisms like Homo‐sapiens (humans), M us, musculus (mouse) and Gallus gallus (chicken) (NCBI). Recently the ICER sequence had been continuously expressed in Danio rerio (zebra fish) through use of the vectors pFlag and TOPO. ICER, which is regulated by the signal path of cAMP, represses the expression of CREM gene is its isoforms. This fact proves to be a favorable influence in the regulation of cancer. Using zebrafish as models, we utilized the C lustered R egular I nterspersed S hort P alindromic R epeats (CRISPR) system with the Cas9 in order to knock out the ICER promoter gene. These were done in two cell lines, embryo fibroblast and tailfin. Our assay results showed the disappearance of the ICER gene, and we will do further experimentation on our cell lines.