We report the nanofabrication and characterization of x-ray transmission gratings with a high aspect ratio and a feature size of down to 65 nm. Two nanofabrication methods, the combination of electron beam and optical lithography and the combination of electron beam, x-ray, and optical lithography, are presented in detail. In the former approach, the proximity effect of electron beam lithography based on a thin membrane of low material was investigated, and the x-ray transmission gratings with a line density of up to 6666 lines/mm were demonstrated. In the latter approach, which is suitable for low volume production, we investigated the x-ray mask pattern correction during the electron beam lithography process and the diffraction effect between the mask and wafer during the x-ray lithography process, and we demonstrated the precise control ability of line width and vertical side-wall profile. A large number of x-ray transmission gratings with a line density of 5000 lines/mm and Au absorber thickness of up to 580 nm were fabricated. The optical characterization results of the fabricated x-ray transmission gratings were given, suggesting that these two reliable approaches also promote the development of x-ray diffractive optical elements.