DNA pol θ Lentiviral Activation Particles (h): sc-407414-LAC

POLQ encodes DNA polymerase theta (DNA pol θ), an error-prone A-family polymerase with an N-terminal helicase-like domain that coordinates end joining and limited DNA synthesis at sites of DNA damage. DNA pol θ is a central component of polymerase theta–mediated end joining (TMEJ), a microhomology-dependent repair pathway that resolves double-strand breaks and promotes genome stability when homologous recombination is compromised. Through its activity in alternative end joining, POLQ influences replication stress tolerance, chromosomal rearrangements, and mutational signatures associated with genomic instability. Dysregulated POLQ activity is frequently studied in the context of cancer-associated DNA repair rewiring and mechanisms of resistance to genotoxic stress, making it relevant for functional genomics of DNA damage response pathways.

DNA pol θ Lentiviral Activation Particles (h) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient POLQ upregulation across a broader range of human cell types.

DNA pol θ Lentiviral Activation Particles (h) deliver all functional components of the synergistic activation mediator (SAM) system via lentiviral transduction. The system comprises three particle preparations co-transduced into target cells: one encoding catalytically inactive dCas9 (D10A and N863A mutations) fused to the VP64 transactivation domain with a blasticidin resistance gene; one encoding the MS2-p65-HSF1 fusion protein with a hygromycin resistance gene; and one encoding a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers with a puromycin resistance gene. Following lentiviral transduction and genomic integration of the expression cassettes, the SAM components are stably expressed and assemble at the target locus within the proximal promoter region upstream of the POLQ transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous DNA pol θ expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native POLQ genomic locus and regulatory architecture.

The lentiviral format offers several practical advantages: stable genomic integration supports heritable activation across cell divisions; high-titer particle preparations eliminate the need for in-house viral production; and compatibility with primary, non-dividing, and transfection-resistant cell types expands experimental accessibility. Successful transduction can be confirmed and enriched through triple antibiotic selection using puromycin, hygromycin, and blasticidin.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.