The KNOP1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of human KNOP1. This heterogeneous pool of HEK293T cells harbors diverse disruptions in the KNOP1 locus, generated by CRISPR/Cas9-mediated gene targeting. The polyclonal format avoids clonal selection, providing a robust model for population-based analyses of ribosome biogenesis and nucleolar biology.
HEK293T is an epithelial cell line derived from human embryonic kidney HEK293 cells, transformed with adenovirus type 5 DNA and stably expressing SV40 large T antigen. This background confers high transfectability and supports episomal plasmid replication, making it a standard host for viral packaging, protein expression, and transient genetic manipulation. The combination of KNOP1 knockout with this versatile cell system offers a flexible platform for studying nucleolar function.
KNOP1 encodes a nucleolar protein essential for ribosome biogenesis. It physically interacts with NOP56, fibrillarin, NPM1, and the U3 snoRNP complex, participating in pre-rRNA processing. KNOP1 promotes maturation of 18S and 28S rRNAs and ribosomal subunit assembly. Its expression is regulated by the MYC transcription factor and mTOR signaling, placing KNOP1 at the nexus of growth control and protein synthesis. Loss of KNOP1 disrupts ribosome production, potentially triggering nucleolar stress and p53-dependent growth arrest.
In HEK293T cells, KNOP1 disruption provides a model to examine ribosome biogenesis defects and nucleolar stress responses. The high transfection efficiency enables rescue studies with exogenous KNOP1 variants, facilitating structure-function analysis. The well-characterized signaling environment permits investigation of MYC- and mTOR-dependent regulation of rRNA processing. This system is also valuable for dissecting the p53-mediated checkpoint activated by nucleolar dysfunction.
Applications include screening ribosome biogenesis inhibitors, validating targets that induce nucleolar stress, and studying cancer cell dependence on increased ribosome production. Researchers can employ immunoblotting, RT-qPCR for rRNA intermediates, immunofluorescence for nucleolar integrity, ribosome profiling, and cell proliferation or apoptosis assays. This ready-to-use polyclonal knockout product accelerates research into nucleolar roles in disease. For further information, contact Ascent Research.
