GENETIC ANALYSIS OF THE INTERRENAL- AND VMH-SPECIFIC ENHANCERS OF THE ZEBRAFISH FF1B GENE

Abstract

IN THIS STUDY, WE HAVE UTILIZED A DELETION BASED APPROACH TO DELETE SPECIFIC INTRONS FROM THE ZEBRAFISH FF1B GENE AND HAVE SUCCESSFULLY LOCALIZED THE PRESENCE OF INTERRENAL GLAND SPECIFIC ENHANCERS IN INTRON IV AND THE VMH-SPECIFIC ENHANCERS IN INTRON V AND VII OF FF1B. FURTHER FRAGMENTATION AND SUB-FRAGMENTATION OF THE FF1B INTRON IV HAS HELPED IDENTIFY A 116 BP INTRON IV FRAGMENT THAT COULD SUCCESSFULLY RECAPITULATE REPORTER EXPRESSION IN THE INTERRENAL GLAND. WE HAVE ALSO TESTED TO CHECK IF THE HUMAN VMH ENHANCER (VMHE) COULD INDUCE MCHERRY REPORTER EXPRESSION IN THE ZEBRAFISH. THE VMHE SUCCESSFULLY RECAPITULATED MCHERRY EXPRESSION IN THE ZEBRAFISH VMH INDICATING THE USE OF SIMILAR TRANSCRIPTIONAL MECHANISMS BETWEEN ZEBRAFISH FF1B AND MAMMALIAN SF. WE THEN DISSECTED THE VMHE IN ORDER TO IDENTIFY SPECIFIC TRANSCRIPTION FACTORS DRIVING THE EXPRESSION TO THE ZEBRAFISH VMH.