The SpaceX Inspiration4 mission was a 3-day mission with four private astronauts to low Earth orbit that occurred in September 2021. The crew collected biospecimen samples before, during, and after flight. One of these biospecimen collections included oral, nasal, and skin microbial swabs collected from ten locations (oral, nasal cavity, post-auricular, axillary vault, volar forearm, occiput, umbilicus, gluteal crease, glabella, and toe web space). Skin swabs were first dipped in nuclease-free water before swabbing each region for 30 seconds. Oral and nasal swabs did not include this swab-wetting step. Nuclease-free water samples were sequenced as controls. Swabs were collected at three pre-flight timespoints (L-92, L-44, L-3), two in-flight timepoints (flight day 2 (FD2), and flight day 3 (FD3)), and three post-flight timepoints (R+1, R+45, R+82). Swabs were stored in 400uL of Zymo Research DNA/RNA Shield at 4C until nucleic acid extraction. DNA and RNA were extracted from the same swab to generate metagenomic and metatranscriptomic profiles.