TALE-SET Fusion Protein Construction

The FokI domain of the TALE-FokI expression vector (pCS2TAL3-DD, Addgene) was replaced with the SET catalytic domain of EHMT2 using the restriction enzymes BamHI and XbaI such that SET was fused to the C-terminal end of the TAL effector. TALE repeat arrays were designed and assembled via multi-fragment cut-ligation using Golden Gate cloning [27 (link)]. These assembled DNA fragments were cloned into the TSET expression vector and the construct was verified by restriction enzyme digestion and sequencing. A 40-amino acid ((GGGS)₁₀) linker was inserted between the TAL effector repeats and the SET domain.

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Cho H.S., Kang J.G., Lee J.H., Lee J.J., Jeon S.K., Ko J.H., Kim D.S., Park K.H., Kim Y.S, & Kim N.S. (2015). Direct regulation of E-cadherin by targeted histone methylation of TALE-SET fusion protein in cancer cells. Oncotarget, 6(27), 23837-23844.

Publication 2015

pCS2TAL3-DD

Amino acid Catalytic Catalytic domain Digestion Ligation Restriction enzyme Restriction enzymes bamhi Set domain Tal effector Vector

Corresponding Organization :

Other organizations : Korea Research Institute of Bioscience and Biotechnology, Korea University of Science and Technology

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Variable analysis

independent variables

Replacement of the FokI domain of the TALE-FokI expression vector (pCS2TAL3-DD, Addgene) with the SET catalytic domain of EHMT2 using the restriction enzymes BamHI and XbaI
Design and assembly of TALE repeat arrays via multi-fragment cut-ligation using Golden Gate cloning

dependent variables

Not explicitly mentioned

control variables

Insertion of a 40-amino acid ((GGGS)10) linker between the TAL effector repeats and the SET domain

controls

No positive or negative controls were explicitly mentioned in the provided information

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