FUS3 transcription factor is a key regulator of seed oil synthesis in plants. In order to investigate the role of the CsFUS3 gene in lipid synthesis and accumulation of Camelina sativa, we conducted a genome-wide identification of the CsFUS3 gene family, analyzed the spatio-temporal expression pattern of CsFUS3 genes, and analyzed the function of CsFUS3-1and CsFUS3-2 genes in lipid synthesis. The results showed: (1) Two CsFUS3 proteins had similar physicochemical properties and high-level structures as Arabidopsis AtFUS3, both had a complete B3 functional domain, and CsFUS3 was most closely related to Arabidopsis AtFUS3 protein. (2) qRT-PCR results indicated that the expression trend of the CsFUS3 genes in seeds increased first and then decreased. (3) The interaction of CsFUS3 and CsWRI1 proteins and the transcriptional regulation of OLE and ABI3 genes by CsFUS3 may be key regulatory pathways of high oil traits in Camelina sativa. (4) Transient expression assays of tobacco showed that CsFUS3 genes could increase the synthesis and accumulation of total oil in tobacco leaves. This study provides a theoretical foundation for in-depth analysis of the function of CsFUS3 genes in the oil synthesis of and the breeding of high oil varieties of Camelina sativa.