The main topic in this article is to define and examine new sequence spaces bs(F(s, r)) and cs(F(s, r))), whereF(s, r) is generalized difference Fibonacci matrix in which s, r ∈ R\ {0}. Some algebric properties including some inclusion relations, linearly isomorphism and norms defined over them are given. In addition, it is shown that they are Banach spaces. Finally, the α-, β-and γ-duals of the spaces bs(F(s, r)) and cs(F(s, r)) are appointed and some matrix transformations of them are given.