Inhibition method of SETDB1 HMTase expression

TALEN is a new concept of cloning genes that can be targeted to a specific gene knockout. TALEN plasmid, since it is a fused DNA binding domain and a Fok1 cutting enzyme function, may be bonded at any one portion of the genomic DNA, the target sequences can induce gene mutations by cutting. The inventors have produced a TALEN structure of two to target the protein initiation codon and upstream promoter -25 region of the gene SETDB1 HMTase. To this end, we conducted a two-step cloning. First, it was confirmed that the target sequence was transferred to the vector pFUS arranged in a module vector, containing approximately 1kb of the insert using the band and the restriction enzyme Esp1 stained via colony PCR. The second was in progress the process of moving to TALEN expression vector from the array vector was confirmed by sequencing. As a result, it was confirmed that the first module is designed vector sequence are arranged at about 100 bp intervals. The fabricated TALEN-DBEX2 structure was confirmed that it was confirmed the disappearance of the expression of SETDB1 via transfection, hayeoteum a mutation occurred in the target site through the T7E1 analysis. On the other hand, through the design TALEN-DBPR25 transfection was confirmed a phenomenon that reduces the expression SETDB1.