ATRX Knockout HEK293T Cell Line

The choice depends on whether you are studying ATRX (α-thalassemia/mental retardation syndrome X-linked)'s role as a chromatin remodeler and H3.3 chaperone or modeling its functions in alternative lengthening of telomeres (ALT) and emerging cancer applications. The Knockout line is the standard tool for asking whether ATRX is required for these processes — ATRX is a SWI/SNF-family chromatin remodeling ATPase that, together with the histone chaperone DAXX, deposits H3.3 at heterochromatin (telomeres, pericentromeres, and other repeats); ATRX maintains telomere stability and represses ALT-mediated telomere maintenance, with critical roles in heterochromatin biology. Overexpression is useful for studying ATRX in heterologous expression contexts. For cancer biology research, the EDITGENE ATRX Knockout in HEK293T is uniquely valuable — HEK293T supports systematic structure-function studies of this large multifunctional protein. ATRX mutations cause ⭐ ATR-X syndrome (X-linked intellectual disability with α-thalassemia, hypogenitalism, characteristic facies); ATRX loss-of-function mutations are characteristic of ⭐⭐ adult diffuse gliomas (especially IDH-mutant astrocytomas) and pancreatic neuroendocrine tumors (panNETs), where ATRX loss enables ALT for telomere maintenance. Rescue with wild-type or ATPase-dead ATRX enables structure-function studies. The knockout is valuable for studying ALT biology, ATR-X syndrome modeling, and emerging ALT-pathway-targeted cancer therapeutics.

Primary applications: • ALT biology: telomere length heterogeneity (alternative lengthening of telomeres) analysis by Southern blot and C-circle assays in ATRX-null cells given ATRX loss enables ALT. • H3.3 deposition: H3.3 ChIP-seq analysis at telomeres and heterochromatin given ATRX-DAXX role. • Pediatric glioma modeling: in heterologous IDH-mutant astrocytoma-relevant contexts, ATRX loss biology characterization. • ATR-X syndrome: rescue with patient-derived ATRX mutations for X-linked intellectual disability disease modeling. • ALT-targeted therapy: ATR inhibitor (e.g., berzosertib, ceralasertib) hypersensitivity in ATRX-null/ALT-positive cells — emerging synthetic lethal approach. EDITGENE recommends this HEK293T-based model for researchers investigating ALT pathway biology and emerging ALT-targeted cancer therapeutics.

Yes, with technical considerations for this large multi-domain protein: • Construct design: use a codon-modified ATRX sequence with a small C-terminal tag (FLAG, HA). ATRX is a ~280 kDa protein with N-terminal ADD (ATRX-DNMT3-DNMT3L) domain and C-terminal SNF2 ATPase domain — full-length rescue is technically challenging due to ~7.5 kb mRNA size. • Domain-focused rescue: ADD or SNF2 domain-focused constructs may be more tractable for specific functional rescue. • ATPase-dead rescue: K1538I in the Walker A motif abolishes ATPase/chromatin remodeling activity. • DAXX partnership: rescue interpretation considers DAXX expression for H3.3 deposition. • Functional readout: rescue should restore ALT suppression and proper H3.3 deposition at telomeres/heterochromatin. HEK293T transduces with very high efficiency and supports stable rescue line generation.