Artemis CRISPR Activation Plasmid (h2): sc-405295-ACT-2

Human DCLRE1C encodes Artemis, a DNA nuclease essential for repairing DNA double-strand breaks during V(D)J recombination and non-homologous end joining, where it cooperates with the DNA-PK complex to open hairpin coding ends and process damaged termini. Artemis activity supports genome stability and proper lymphocyte development by enabling accurate antigen receptor gene assembly and resolving complex DNA lesions generated by ionizing radiation or replication stress. Loss-of-function variants in DCLRE1C are linked to severe combined immunodeficiency and radiosensitivity, highlighting its role at the intersection of adaptive immunity and DNA damage response signaling. Gene editing and functional genomics studies targeting DCLRE1C/Artemis are used to model NHEJ pathway defects, interrogate end-processing requirements in double-strand break repair, and evaluate impacts on chromosomal rearrangements and immune repertoire formation in human cells.

Artemis CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous DCLRE1C expression without altering the underlying DNA sequence.

Artemis CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DCLRE1C locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the DCLRE1C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Artemis expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DCLRE1C locus and enabling the study of Artemis-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Artemis pathway restoration in tumor cells with silenced or reduced DCLRE1C expression.

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