Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
TdT CRISPR/Cas9 KO Plasmid (m): sc-423324
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TdT CRISPR/Cas9 Knockout (KO) Plasmid (m) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the TdT genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TdT Antibody (C-11): sc-393710
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TdT CRISPR/Cas9 KO Plasmid (m) | sc-423324 | 20 µg | $397.00 |
Overview
DNA nucleotidylexotransferase (Dntt) encodes terminal deoxynucleotidyl transferase (TdT), a template-independent DNA polymerase that adds non-templated nucleotides to V(D)J recombination junctions to generate N-region diversity. TdT functions in developing lymphocytes during antigen receptor gene assembly, influencing repertoire breadth through modulation of end-joining outcomes and junctional processing. Its activity intersects with DNA double-strand break repair and non-homologous end joining (NHEJ) factors that resolve recombination intermediates. Dysregulated Dntt expression or aberrant TdT activity is widely used as a molecular feature of immature lymphoid states and is relevant for mechanistic studies of lymphoid development and genome integrity during recombination.
TdT CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Dntt gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Dntt together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the Dntt open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish TdT protein expression.
This CRISPR knockout system enables efficient generation of Dntt-deficient cell models for investigation of TdT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Dntt exon(s) critical for TdT function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple Dntt genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TdT CRISPR/Cas9 KO Plasmid (m) and TdT CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Dntt locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TdT HDR Plasmid (m) and TdT HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Dntt homology arms to support homology-directed repair at defined Dntt target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.