Chemical vapor deposition of (111) 3C-SiC on (110) Si substrate was carried out, and the effect of the substrate off-axis introduced on (110) Si substrate for suppressing the twin formation in 3C-SiC hetero-epitaxial layers was investigated. From the growth on hemispherically polished (110) Si substrate, it was found that the off-axis toward the [001]-Si axis had a noble effect for suppressing the twin formation, while the off-axis toward the [110] Si axis was ineffective. The growth of single 3C-SiC crystal containing few double positioning boundaries, which are related with the twin formation, was demonstrated on the (110) Si substrate 3° off-axis toward the [001] Si axis. Transmission electron microscopic observation revealed that double positioning boundaries on the (110) Si substrate off-axis toward the [001] Si axis were nearly eliminated within the initial a few hundreds nano meter in thickness.