IMPC phenotypes (gene matching)
C57BL/6-Rag2em1Bdes Csf1em1(CSF1)Bdes Appem1Bdes/Ph
| Status | Only small colony available |
| EMMA ID | EM:15929 |
| Citation information | RRID:IMSR_EM:15929 Research Resource Identifiers (RRID) are persistent unique ID numbers assigned to help researchers cite key resources (e.g. antibodies, model organisms and software projects) in the biomedical literature to improve transparency and reproducibility in research. See https://www.rrids.org/ for more information. |
| International strain name | C57BL/6-Rag2em1Bdes Csf1em1(CSF1)Bdes Appem1Bdes/Ph |
| Alternative name | Csf1em1(CSF1)Bdes;Rag2em1Bdes;Appem1Bdes - short name hCSF1Bdes;Apphu |
| Strain type | Endonuclease-mediated |
| Allele/Transgene symbol | Rag2em1Bdes, Appem1Bdes, Csf1em1(CSF1)Bdes |
| Gene/Transgene symbol | Rag2, App, Csf1 |
Information from provider
| Provider | Bart De Strooper |
| Provider affiliation | KU Leuven and VIB |
| Additional owner | This mouse strain was produced in collaboration between University College London and KULeuven. |
| Genetic information | Humanisation of the Csf1 gene by endonuclease mediated gene targeting. Knock-out of the Rag2 gene by endonuclease mediated gene targeting. Humanisation of the Abeta sequence in App gene by endonuclease mediated gene targeting. The hCSF1Bdes;Apphu triple-mutant mouse strain facilitates the use of the xenotransplantation paradigm for human microglia to study complex human disease. The humanization of Abeta sequence in the AppHu model makes this the control model to use to study Alzheimer disease. |
| Phenotypic information | Homozygous:Mice are lacking T and B cells (immune deficient) and express human CSF1, which make make them suitable for xenografting human microglia. Mice have a humanized Abeta sequence in the App gene, which makes that they produce ~3-fold more Abeta compared to mouse.Heterozygous:no abnormal phenotype |
| Breeding history | CRISPR/Cas9 technology was used to replace the genomic DNA sequence corresponding to amino acid 33–552 of the mouse Csf1 gene with the human CSF1 genomic sequence corresponding to amino acid 33–554 in the endogenous gene. The 5’ murine signal peptide (amino acid 1–32) and 3’ region of the mouse were retained. The gRNAs (target sequence ACA GTG TTC TGA CAC CTC CTTGG and GTG GAA CTG CCA GTA TAG AAAGG ) to the mouse Csf1 gene, the donor vector (PCR assembled from BAC: RP23-15F10 (mouse) and BAC: RP11-101M23 (human)), and Cas9 mRNA were co-injected into fertilized mouse eggs of C57BL/6J mice. F0 founder animals were identified by extensive PCR analysis followed by Sanger sequence analysis. F0 were bred to wild-type mice to test germline transmission and generate F1 animals. F1 animals were backcrossed to wild-type mice to generate F2. This line is called Csf1em1(CSF1)Bdes. Rag2em1Bdes knock-out: CRISPR/Cas9 technology (target sequence AAC ATA GCC TTA ATT CAA CCAGG and ACG AGT CTG TAA CCG GCT ACTGG and Cas9 enzyme obtained from IDT) was used to generate an F0 founder in C57BL/6J mice. The F0 founder was selected by extensive analysis utilizing PCR and sequencing which demonstrated a 1198 bp genomic DNA deletion which corresponds to amino acid 16–430 and generates a premature stop. The F0 founder was backcrossed two times to wild-type mice. Mice Appem1Bdes with a humanized Abeta sequence (G676R, F681Y, R684H) were generated using CRISPR/Cas9 technology to target exon 16 of the mouse App gene. RNA guides were selected using the CRISPOR web tool. Guide 5′-GCAGAAUUCGGACAUGAUUC-3′ and 5′-GUCCGCCAUCAAAAACUGGU-3′ were selected and tested in mouse embryonic fibroblast (MEF) cells for cleaving efficiency. To promote homologous recombination directed repair we made use of a ssODN repair template to mutate the target amino acids and to introduce two silent nucleotide substitutions. The first silent substitution destroys an EcoRI restriction cleaving site, facilitating genotyping. The second silent substitution prevents Cas9 cleaving the modified locus. Ribonucleoproteins (RNPs) containing 0.3 μM purifiedCas9HiFi protein (Integrated DNA Technologies,IDT), 0.6 μM CRISPR RNAcrRNA, 0.6 μM trans-activating crRNA (IDT) and 10 ng/μl ssODN (5′ tactttgtgtttgacg cagGTTCTGGGCTGACAAACATCAAGACGGAAGA GATCTCGGAAGTGAAGATGGATGCAGAATTtaGA CATGATTCAGGATaTGAAGTCCaCCATCA gAAACTGgtaggcaaaaataaactgcctctccccgagattgcgtctggc cagatgaaatacgtggcacctcgtggcttgtcctgtgt-3′) were injected into the pronucleus of 72 C57BL/6J embryos by microinjection in the Mouse Expertise Unit of KU Leuven. One positive pup was identified by PCR and restriction analysis. Sanger sequencing of App exon 16 region, as well as the 5 most likely off target sites predicted by the CRISPOR web tool, confirmed correct targeting and absence of spurious events at other sites. The founder mouse was backcrossed over two generations using C57BL/6J mice before a homozygous colony was established, which was designated Apphu/hu. This strain was crossed with the Csf1em1(CSF1)Bdes;Rag2em1Bdes strain (EMMA strain EM:15928). The short name of the triple-mutant strain is hCSF1Bdes;Apphu. The three gene mutations are kept homozygous. |
| References |
|
| Homozygous fertile | yes |
| Homozygous viable | yes |
| Homozygous matings required | yes |
| Immunocompromised | yes |
Information from EMMA
| Archiving centre | Institute of Molecular Genetics, Prague, Czech Republic |
Disease and phenotype information
Orphanet associated rare diseases, based on orthologous gene matching
- ABeta amyloidosis, Italian type / Orphanet_324713
- ABetaL34V amyloidosis / Orphanet_324703
- ABeta amyloidosis, Arctic type / Orphanet_324723
- ABetaA21G amyloidosis / Orphanet_324718
- ABeta amyloidosis, Dutch type / Orphanet_100006
- ABeta amyloidosis, Iowa type / Orphanet_324708
- Early-onset autosomal dominant Alzheimer disease / Orphanet_1020
- Severe combined immunodeficiency due to complete RAG1/2 deficiency / Orphanet_331206
- Omenn syndrome / Orphanet_39041
- Combined immunodeficiency with granulomatosis / Orphanet_157949
MGI phenotypes (gene matching)
- amyloidosis / MGI
- muscle degeneration / MGI
- decreased corpus callosum size / MGI
- abnormal telencephalon morphology / MGI
- decreased body weight / MGI
- increased anxiety-related response / MGI
- abnormal locomotor behavior / MGI
- hyperactivity / MGI
- hypoactivity / MGI
- decreased exploration in new environment / MGI
- abnormal cued conditioning behavior / MGI
- abnormal spatial learning / MGI
- reduced long term potentiation / MGI
- impaired swimming / MGI
- intracerebral hemorrhage / MGI
- abnormal learning/memory/conditioning / MGI
- abnormal fear/anxiety-related behavior / MGI
- abnormal motor capabilities/coordination/movement / MGI
- premature death / MGI
- abnormal brain morphology / MGI
- no abnormal phenotype detected / MGI
- decreased brain weight / MGI
- gliosis / MGI
- absent corpus callosum / MGI
- abnormal brain commissure morphology / MGI
- abnormal long term potentiation / MGI
- abnormal emotion/affect behavior / MGI
- increased thigmotaxis / MGI
- abnormal active avoidance behavior / MGI
- abnormal long term object recognition memory / MGI
- abnormal neuron morphology / MGI
- no phenotypic analysis / MGI
- abnormal locomotor activation / MGI
- amyloid beta deposits / MGI
- astrocytosis / MGI
- nervous system phenotype / MGI
- impaired passive avoidance behavior / MGI
- myositis / MGI
- homeostasis/metabolism phenotype / MGI
- growth/size/body region phenotype / MGI
- behavior/neurological phenotype / MGI
- abnormal cell physiology / MGI
- decreased hippocampal commissure size / MGI
- decreased anterior commissure size / MGI
- abnormal spatial reference memory / MGI
- hematoma / MGI
- microgliosis / MGI
- increased variability of skeletal muscle fiber size / MGI
- centrally nucleated skeletal muscle fibers / MGI
- abnormal synapse morphology / MGI
- decreased grip strength / MGI
- altered susceptibility to induced thrombosis / MGI
- abnormal circadian behavior / MGI
- fragile skeleton / MGI
- abnormal bone marrow cavity morphology / MGI
- osteoporosis / MGI
- osteopetrosis / MGI
- abnormal parietal bone morphology / MGI
- failure of tooth eruption / MGI
- absent incisors / MGI
- abnormal trabecular bone morphology / MGI
- decreased compact bone thickness / MGI
- abnormal microglial cell morphology / MGI
- abnormal long bone hypertrophic chondrocyte zone / MGI
- decreased circulating phosphate level / MGI
- decreased leukocyte cell number / MGI
- decreased monocyte cell number / MGI
- extramedullary hematopoiesis / MGI
- increased granulocyte number / MGI
- decreased bone marrow cell number / MGI
- domed cranium / MGI
- short limbs / MGI
- abnormal tibia morphology / MGI
- abnormal femur morphology / MGI
- abnormal autopod morphology / MGI
- short tail / MGI
- abnormal mammary gland development / MGI
- abnormal branching of the mammary ductal tree / MGI
- small spleen / MGI
- thymus cortex hypoplasia / MGI
- abnormal ovary morphology / MGI
- abnormal ovarian folliculogenesis / MGI
- decreased body weight / MGI
- abnormal mating frequency / MGI
- reduced male mating frequency / MGI
- abnormal eating behavior / MGI
- abnormal circulating calcium level / MGI
- impaired embryo implantation / MGI
- abnormal postnatal growth / MGI
- postnatal growth retardation / MGI
- abnormal lactation / MGI
- reduced fertility / MGI
- reduced male fertility / MGI
- reduced female fertility / MGI
- abnormal ovulation / MGI
- decreased litter size / MGI
- premature death / MGI
- abnormal spleen red pulp morphology / MGI
- abnormal bone marrow morphology / MGI
- abnormal bone marrow cell morphology/development / MGI
- abnormal sperm number / MGI
- decreased corpora lutea number / MGI
- decreased mature ovarian follicle number / MGI
- oligozoospermia / MGI
- decreased circulating testosterone level / MGI
- abnormal bone remodeling / MGI
- no phenotypic analysis / MGI
- amyloid beta deposits / MGI
- decreased ovulation rate / MGI
- impaired granulosa cell differentiation / MGI
- increased width of hypertrophic chondrocyte zone / MGI
- abnormal uterus development / MGI
- abnormal bone structure / MGI
- abnormal compact bone morphology / MGI
- impaired macrophage chemotaxis / MGI
- decreased macrophage cell number / MGI
- abnormal masseter muscle morphology / MGI
- decreased length of long bones / MGI
- abnormal thyroid parafollicular C-cell morphology / MGI
- abnormal auditory brainstem response / MGI
- abnormal vestibular system physiology / MGI
- abnormal hematopoietic stem cell morphology / MGI
- increased testis weight / MGI
- abnormal testis physiology / MGI
- decreased spleen weight / MGI
- abnormal osteoclast morphology / MGI
- abnormal osteoclast cell number / MGI
- decreased osteoclast cell number / MGI
- abnormal osteoblast morphology / MGI
- decreased bone resorption / MGI
- abnormal osteoblast physiology / MGI
- increased T cell number / MGI
- decreased lymphocyte cell number / MGI
- decreased B cell number / MGI
- decreased susceptibility to atherosclerosis / MGI
- abnormal Langerhans cell physiology / MGI
- behavior/neurological phenotype / MGI
- osteosclerosis / MGI
- abnormal skeleton morphology / MGI
- increased bone mass / MGI
- abnormal mammary gland growth during pregnancy / MGI
- abnormal mammary gland growth during lactation / MGI
- increased long bone epiphyseal plate size / MGI
- abnormal Kupffer cell morphology / MGI
- abnormal Langerhans cell morphology / MGI
- absent Langerhans cell / MGI
- abnormal perivascular macrophage morphology / MGI
- abnormal myeloid leukocyte morphology / MGI
- abnormal osteoclast differentiation / MGI
- abnormal neurite morphology / MGI
- slow postnatal weight gain / MGI
- abnormal osteocyte morphology / MGI
- abnormal splenic cell ratio / MGI
- abnormal ovarian follicle number / MGI
- decreased physiological sensitivity to xenobiotic / MGI
- prolonged estrous cycle / MGI
- abnormal proestrus / MGI
- myometrium hypoplasia / MGI
- endometrium hypoplasia / MGI
- necrospermia / MGI
- abnormal splenocyte physiology / MGI
- decreased trabecular bone thickness / MGI
- abnormal mammary gland lobule morphology / MGI
- abnormal mammary gland alveolus morphology / MGI
- decreased fertilization frequency / MGI
- decreased hippocampus pyramidal cell number / MGI
- abnormal visual evoked potential / MGI
- increased bone trabecula number / MGI
- increased trabecular bone mass / MGI
- increased trabecular bone volume / MGI
- postnatal lethality, incomplete penetrance / MGI
- decreased secondary ovarian follicle number / MGI
- abnormal tooth root development / MGI
- impaired osteoblast differentiation / MGI
- abnormal auditory brainstem response waveform shape / MGI
- thymus medulla atrophy / MGI
- decreased bone mineralization / MGI
- round snout / MGI
- abnormal dentin mineralization / MGI
- increased osteocyte apoptosis / MGI
- abnormal osteocyte lacuna morphology / MGI
- abnormal osteocyte dendritic process morphology / MGI
- abnormal dental follicle morphology / MGI
- abnormal predentin morphology / MGI
- absent teeth / MGI
- decreased monocyte cell number / MGI
- increased granulocyte number / MGI
- decreased bone marrow cell number / MGI
- alopecia / MGI
- abnormal digestive system morphology / MGI
- abnormal intestinal epithelium morphology / MGI
- abnormal intestinal mucosa morphology / MGI
- abnormal spleen morphology / MGI
- small spleen / MGI
- spleen hypoplasia / MGI
- abnormal Peyer's patch morphology / MGI
- abnormal thymus morphology / MGI
- decreased thymocyte number / MGI
- abnormal immune system cell morphology / MGI
- decreased body size / MGI
- abnormal humoral immune response / MGI
- arrested B cell differentiation / MGI
- decreased IgG level / MGI
- decreased IgM level / MGI
- decreased IgA level / MGI
- thymus hypoplasia / MGI
- arrested T cell differentiation / MGI
- lung inflammation / MGI
- neoplasm / MGI
- abnormal B cell differentiation / MGI
- no abnormal phenotype detected / MGI
- small lymph nodes / MGI
- abnormal lymph node morphology / MGI
- abnormal lymphopoiesis / MGI
- abnormal pre-B cell morphology / MGI
- increased susceptibility to infection / MGI
- abnormal double-negative T cell morphology / MGI
- abnormal double-positive T cell morphology / MGI
- increased susceptibility to bacterial infection / MGI
- abnormal effector T cell morphology / MGI
- abnormal macrophage physiology / MGI
- abnormal B cell number / MGI
- abnormal B cell physiology / MGI
- abnormal immune system organ morphology / MGI
- colitis / MGI
- absent Peyer's patches / MGI
- increased natural killer cell mediated cytotoxicity / MGI
- erythroderma / MGI
- abnormal lymphocyte physiology / MGI
- abnormal B cell morphology / MGI
- decreased lymphocyte cell number / MGI
- decreased B cell number / MGI
- decreased T cell number / MGI
- decreased susceptibility to parasitic infection / MGI
- decreased double-negative T cell number / MGI
- increased double-negative T cell number / MGI
- decreased double-positive T cell number / MGI
- decreased T cell proliferation / MGI
- cachexia / MGI
- increased macrophage cell number / MGI
- abnormal T-helper 1 physiology / MGI
- decreased susceptibility to type IV hypersensitivity reaction / MGI
- abnormal response to transplant / MGI
- abnormal T cell morphology / MGI
- decreased NK T cell number / MGI
- decreased CD4-positive, alpha beta T cell number / MGI
- decreased CD8-positive, alpha-beta T cell number / MGI
- lymph node hypoplasia / MGI
- decreased pre-B cell number / MGI
- decreased mature B cell number / MGI
- absent mature B cells / MGI
- increased immature B cell number / MGI
- decreased immature B cell number / MGI
- abnormal gamma-delta T cell differentiation / MGI
- decreased spleen white pulp amount / MGI
- abnormal interferon secretion / MGI
- abnormal chemokine secretion / MGI
- abnormal T cell receptor V(D)J recombination / MGI
- abnormal immunoglobulin V(D)J recombination / MGI
- absent Hassall's corpuscle / MGI
- small inguinal lymph nodes / MGI
- decreased susceptibility to bacterial infection induced morbidity/mortality / MGI
- increased DN3 thymocyte number / MGI
- abnormal stomach mucosa morphology / MGI
- small cervical lymph nodes / MGI
Literature references
- A versatile mouse model to advance human microglia transplantation research in neurodegenerative diseases.;Serneels Lutgarde, Sierksma Annerieke, Pasciuto Emanuela, Geric Ivana, Nair Arya, Martinez-Muriana Anna, Snellinx An, De Strooper Bart, ;2025;Molecular neurodegeneration;20;29; 40069774
- Modeling the β-secretase cleavage site and humanizing amyloid-beta precursor protein in rat and mouse to study Alzheimer's disease.;Serneels Lutgarde, T'Syen Dries, Perez-Benito Laura, Theys Tom, Holt Matthew G, De Strooper Bart, ;2020;Molecular neurodegeneration;15;60; 33076948