Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease characterized by loss of upper and lower motor neurons, leading to paralysis and death within 3–5 years after diagnosis. Mutations in the C-terminus of human KIF5A (kinesin-1 family member 5A) were identified as the genetic cause of ALS and reported to have high penetrance. KIF5A encodes for a motor protein involved in cargo trafficking such as cell organelles, proteins, RNA, and lipids. Based on our recent published data, the gain-of-function (GOF) hypothesis proposed that the KIF5A ALS mutant is hyperactive, increased motor self-association, aggregate formation, and enhanced processivity on microtubules – all correlating with premature cell death. However, the molecular mechanism of KIF5A-associated ALS is poorly understood, and it is not currently known how this GOF behavior of KIF5A causes disease. To understand this, we generated a novel transgenic mouse model that expresses human KIF5A (wild-type or ALS form) under the control of neuronal-specific promoter. The expression of human transgene is comparable to endogenous mouse Kif5a with a minimal concern of high overexpression. We are currently examining these mice in a variety of behavioral, neurophysiological, histological, and molecular levels. The full characterization of these mice will provide an additional model for preclinical studies in ALS.