The DTWD1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DTWD1 gene in the human embryonic kidney HEK293T cell line. This heterogeneous pool of edited cells provides a loss-of-function model for investigating the role of DTWD1 in tRNA modification and translation regulation.
The HEK293T cell line is a widely utilized epithelial cell model derived from human embryonic kidney, transformed with the SV40 large T antigen and adenovirus 5. Its robust growth, high transfection efficiency, and genetic tractability make it an ideal host for generating knockout models and performing downstream mechanistic studies.
DTWD1 encodes a probable tRNA (cytidine/uridine-2′-O-)-methyltransferase that catalyzes 2′-O-methylation of specific tRNA residues, utilizing S-adenosylmethionine (SAM) as a methyl donor. This modification is critical for tRNA stability, folding, and efficient translation. DTWD1 operates within the tRNA maturation pathway, likely interacting with components of the tRNA methyltransferase complex such as TRMT61A and TRMT6. Its activity modulates the translational machinery by influencing tRNA substrate recognition, thereby impacting protein synthesis fidelity and cellular stress responses.
In HEK293T cells, disruption of DTWD1 is anticipated to alter tRNA modification profiles, leading to changes in global protein synthesis and possibly activation of stress pathways. Given the role of tRNA modifications in regulating translation under physiological and stress conditions, this knockout model enables the study of how aberrant tRNA methylation affects translational control, cell proliferation, and potentially oncogenic processes. The polyclonal nature of the population preserves heterogeneity, allowing the observation of diverse editing outcomes and functional consequences.
Researchers can employ these cells in epitranscriptomic studies to dissect the impact of tRNA 2′-O-methylation on gene expression. Representative assays include RiboMeth-seq for comprehensive tRNA methylation profiling, ribosome profiling to assess translational efficiency, and Western blotting to confirm DTWD1 loss. Proliferation and viability assays can be used to examine phenotypic effects. Validation of gene disruption can be performed via Sanger sequencing and RT-qPCR. For further information or to request a quote, please contact Ascent Research.
