The Human Silencing Hub (HuSH) complex is a complex that silences retrotransposable elements in
vertebrates. Here, we identify a second HuSH complex, designated HuSH2, which is centered around
TASOR2, a paralog of the core TASOR protein in HuSH. Our findings reveal that HuSH and HuSH2 localize to
distinct and non-overlapping genomic loci. Specifically, HuSH localizes to and represses LINE-1
retrotransposons, whereas HuSH2 targets and represses KRAB-ZNFs and interferon signaling and response
genes. We use in silico protein structure predictions to simulate MPP8 interactions with TASOR paralogs,
guiding amino acid substitutions that disrupted binding to HuSH complexes. These MPP8 transgenes and
other constructs reveal the importance of HuSH complex quantities in regulating LINE-1 activity. Furthermore,
our results suggest that dynamic changes in TASOR and TASOR2 expression enable cells to finely tune
HuSH-mediated silencing. This study offers insights into the interplay of HuSH complexes, highlighting their
vital role in retrotransposon regulation.