N2G mouse
The N2G mouse (for NOD–2-Genes knockout) is a severely immunodeficient laboratory mouse strain established on a NOD background. It was generated using CRISPR/Cas9 technology to delete nearly the entire genomic regions of both Prkdc (DNA-PKcs) and Il2rg (common γ chain).[1] The model was developed to facilitate human cell and tissue engraftment in oncology and immune-system research, addressing limitations of earlier immunodeficient strains that retained residual mutant transcripts or selection cassettes.[2]
Severe combined immunodeficiency in the widely used NSG and NOG strains results from the combination of a Prkdc mutation and Il2rg deficiency, eliminating mature T and B cells as well as functional natural killer (NK) cells and thereby enabling efficient human xenograft engraftment.[3][4] The N2G strain was developed with the same purpose and exhibits functionally equivalent immunodeficient characteristics, but it was re-created using an improved CRISPR/Cas9-based large-scale deletion strategy that avoids residual transcripts and selection cassettes, providing a refined platform for humanization studies. The strain was developed by GEMCRO Inc. in Korea.[5]
Background
The NSG and NOG strains have become indispensable in translational research for oncology, virology, immunology, and regenerative medicine.[3][4] They support engraftment of human cells and tissues, including hematopoietic stem cells (HSCs), and are widely used for cancer xenograft and immune system reconstitution studies.[6]
Development and genetics
N2G was produced on a NOD background by microinjection of Cas9 protein and guide RNAs into zygotes to excise broad genomic segments: exons 3 through the 3′ UTR of Prkdc and exons 1 through the 3′ UTR of Il2rg.[1] This strategy yielded near-complete deletions that abolished detectable transcripts of both genes in bone marrow, spleen, and thymus.[1]
- Prkdc encodes DNA-PKcs, required for V(D)J recombination; its loss causes a T–B– SCID phenotype.[2]
- Il2rg encodes the common γ chain; null alleles result in NK-cell deficiency and broader lymphoid defects.[3][4]
Immunophenotype
N2G mice lack functional T cells, B cells, and NK cells.[1] Transcript analysis indicates minimal to undetectable Prkdc and Il2rg expression across hematolymphoid tissues, with reduced age-related immune “leakiness”[1]
Applications
Cancer xenografts
N2G supports subcutaneous growth of human cancer cell lines at levels comparable to NOG, making it suitable for CDX and PDX studies.[1]
Humanized immune system (HIS) mice
The strain enables robust engraftment of human hematopoietic stem and progenitor cells, with higher levels of peripheral T-cell reconstitution compared with NSG.[1] Conditioning regimens include non-irradiation alternatives such as busulfan.[1]
PBMC humanization
As in other Il2rg-null NOD strains, PBMC engraftment is feasible but limited by xenogeneic graft-versus-host disease (xeno-GVHD), typically emerging 4–8 weeks after transplantation.[6]
See also
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- ↑ 1.0 1.1 1.2 1.3 1.4 1.5 1.6 1.7 Kim, Y.M.; Na, H.J.; Kwon, D.H. (2025). "Generation of NOD SCID mice with near-complete deletions of Il2rg and Prkdc for human cancer and HSC engraftment". Transgenic Research. 34 (1): 35. doi:10.1007/s11248-025-00454-9. PMC 12254168 Check
|pmc=value (help). PMID 40643795 Check|pmid=value (help). - ↑ 2.0 2.1 Mathieu, A.L. (2013). "PRKDC mutations associated with immunodeficiency in humans and animals". Journal of Clinical Investigation. 123 (9): 4001–4011. doi:10.1172/JCI68943. PMC 4381280. PMID 23979165.
- ↑ 3.0 3.1 3.2 "NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG)". The Jackson Laboratory. 2019. Retrieved 2025-09-10.
- ↑ 4.0 4.1 4.2 "CIEA NOG mouse®". Taconic Biosciences / CIEA. 2024. Retrieved 2025-09-10.
- ↑ "N2G immunodeficient mouse (overview)". GEMCRO Inc. 2025. Retrieved 2025-09-10.[permanent dead link]
- ↑ 6.0 6.1 Cogels, M.M. (2021). "Humanized mice as a valuable pre-clinical model for cancer immunotherapy". Frontiers in Oncology. 11: 784098. doi:10.3389/fendo.2021.777831. PMC 8723090 Check
|pmc=value (help). PMID 34975753 Check|pmid=value (help).
