To track changes in CPSF6 localization following HIV-1 infection, we engineered endogenous monomeric NeonGreen (mNGreen) C-terminal tagged CPSF6 into HT1080 cells (HT1080-CPSF6-mNGreen) via CRISPR-Cas9 knock-in. Here, CPSF6 is linked to HIV-1 infection.