Mouse Gene Editing

We gene edit several different strains of mice, including C57BL/6, Balb/c, DBA2, and NOD-scid for use as disease models. Mouse model generation can use either CRISPR/CAS9 EGE™ or ESC-based gene editing technologies, for which the turnaround times are 6-8 months and 9-12 months, respectively. Mice are often the preferred mammalian model organisms due to their short life cycle and close genetic resemblance to humans. Particularly, humanized mouse models have great potential.  Mice happen to be the best studied mammalian model organism with abundant genomic, transcriptomic, proteomic, and phenotypic collections.

  • Strain: Laboratory mouse strains are either outbred or inbred. Outbred mice are usually bred to maintain maximum heterozygosity, whereas inbred mice are usually bred to maintain maximum homozygosity. Inbred strains (also called inbred lines) are particular species that have individual mice nearly identical to each other in genotype due to long inbreeding. A strain is inbred when it has undergone at least 20 generations of brother x sister or offspring x parent mating, at which point at least 98.6% of the loci in an individual of the strain will be homozygous, and each individual can be treated effectively as clones. C57BL/6(N and J), BALB/c, Fvb, A/J, C3H, CBA, DBA/2 etc. are inbred mouse strains.
  • CRISPR/EGE™ or ESC/HR: At Biocytogen, 85% mouse and 100% rat gene editing projects are generated via EGE™ technology. Although EGE™ technology is comparatively cheaper and offers faster turnaround times compared to ESC-based projects, projects where the knockin size is greater than 15kb still necessitate the use of ESC/HR technology.
  • Superovulation: Superovulation has been used in the production of transgenic mice since the late 1980s to artificially induce ovulation of large numbers of oocytes from limited numbers of female mice. Superovulation facilitates the generation of genetically engineered mice and reduces the number of animals used.
  • Breeding: Breeding is one of the critical steps to generate genetically engineered mouse models. A suitable breeding environment is important to have large litter sizes and shorten the timelines to produce mouse models. Biocytogen’s AAALAC accredited SPF animal facilities provide optimal breeding environments. Our experts design the most efficient breeding scheme to save on costs and time for our customers.

CRISPR Knockin Mouse Application Example:

Humanized CD3ε Mouse Model Generation and Validation

Targeting Strategy

Mouse-Targeting-Strategy

mRNA Expression Analysis

RT-PCR analysis of the B-hCD3ε gene: hCD3ε mRNA was detected in splenocytes isolated from homozygous B-hCD3ε mice.

mRNA-Expression-Analysis

Protein Expression Analysis

Splenocytes from both wild type (WT) C57BL/6 and homozygous B-hCD3ε mice were analyzed by flow cytometry.

Results: Mouse CD3+ cells were detectable in the WT C57BL/6 mice, whereas human CD3+ cells were detectable in the homozygous knockin mouse B-hCD3ε.

Mouse-Protien-Expression-Analysis

Triple Humanized

Product Product No Background
B-hPD-1/hPD-L1/hTIGIT mice 130573 C57BL/6
B-hPD-1/hPD-L1/hTNFR2 mice 130849 C57BL/6
B-hPD1/hLAG3/hTIM3 mice 130999 C57BL/6

Other Genetically Modified Mouse Models

Product Product No Application Fields
B-Apoe-KO Mice B-EM-021 Hyperlipidemia, atherosclerosis mice model (Cardiovascular system)
B-DTR-EGFP cKI Mice 110151 DTR conditional Knock-in mice model
B-F8 KO Mice B-CM-007 Hemophilia mice model
B-F9 KO Mice B-CM-008 Hemophilia mice model
B-Il17a-EGFP Mice B-EM-017 IL17 expression and functional test (gene reporter mice)
B-Il17a-Gluc Mice B-EM-019 IL17 expression and functional test (gene reporter mice)
B-db/db Mice B-CM-012 Diabetes mice model
B-ob/ob Mice B-CM-059 Diabetes mice model
B-p53 KO Mice B-EM-020 Tumorigenesis drug test
TGFb1-cKO Mice B-EM-022 Study on transforming growth factor

Cre Mouse Models List

Product Product No Application Fields
B-ALB-CreERT2 Mice 110137 Alb promoter-driven inducible Cre mouse model (liver)
B-CAG-iCreERT2 mice 110147 Cre ubiquitous expression in mice
B-CAG-tdTomato cKI Mice 110148 Cre-induced tdTomato expression, for Cre expression profile detection
B-CD150-iCreERT2 Mice 110142 CD150 promoter-driven inducible Cre mouse model (Thymic, T cells and macrophages)
B-Cam1(Mcpt5)-iCre mice 110138
B-Cdh5-iCreERT2 mice 110140 Cdh5 promoter-driven inducible Cre mouse model ( lung adult, subcutaneous fat pad adult)
B-Col10a1-iCre Mice 110122 Col10a1 promoter-driven Cre mouse model (mature hypertrophic chondrocyte)
B-Col2a1-iCre Mice 110123 Col2 promoter-driven Cre mouse model (type II collencyte)
B-Dcp2-iCreERT2 mice 110138 Dcp2 promoter-driven inducible Cre mouse model. (Macrophages)
B-Dmp1-iCre Mice 110124 Dmp1 promoter-driven Cre mouse model (skeleton)
B-Gfap-iCre mice 110155 Gfappromoter-driven Cre mouse model. (Astroglia)
B-Grpr-iCre Mice 110125 Grpr promoter-driven Cre mouse model (digestive system, nervous system)
B-Ins1-iCreERT2 Mice 110126 Ins1 promoter-driven inducible Cre mouse model (pancreas isletbeta cell)
B-Myh6-iCreERT2 mice 110127 Myh6 promoter-driven inducible Cre mouse model (Heart)
B-Ncr1 (Nkp46)-iCre Mice 110127 Ncr1 promoter-driven Cre mouse model (NK cell)
B-Oxr1-iCreERT2 Mice 110128 Oxr1 promoter-driven inducible Cre mouse model (frontal lobe adult, cerebellum adult and 24 other tissues)
B-Pdgfrb-iCre Mice 110129 Pdgfrb promoter-driven inducible Cre mouse model (parietal cell of multiple organs, such as skin, intestinal tract, kidney, lung and plexus vasculosus around central nervous system)
B-Prrx1-iCre Mice 110130 Prrx1 promoter-driven Cre mouse model (mesenchyme)
B-Sp7 (Osx)-iCre Mice 110131 Sp7 promoter-driven Cre mouse model (skeleton)
B-Syn1-iCre Mice B-CM-005 Syn1promoter-driven inducible Cre mouse model (brain)
B-Tcf21-iCreERT2 mice 110141 Tcf21 promoter-driven inducible Cre mouse model (bladder, lung)
B-Tex264-iCre-EGFP mice 110136 Tex264 promoter-driven Cre mouse model(Ubiquitous expression in adrenal,kidney and 28 other tissues)
B-Tnfrsf11a (RANK)-iCre Mice B-EM-009 Tnfrsf11a promoter-driven Cre mouse model (lymph gland formation and osteoclast differentiation system)
B-Ucp1-iCre Mice 110134 UCP1 promoter-driven Cre mouse model (mitochondria, brown fat)
B-Zeb1-iCreERT2 Mice 110135 Zeb1 promoter-driven inducible Cre mouse model (tunica media of artery and myocardial cell)

Single Humanized Immune-checkpoint Mice

Product Product No Background
B-hB7-H3 mice 110028 C57BL/6
B-hBTLA mice 110005 C57BL/6
B-hCD27 mice 110006 C57BL/6
B-hCD28 mice 110007 C57BL/6
B-hCD38 mice 110046 C57BL/6
B-hCD3E mice 110008 C57BL/6
B-hCD40 mice 110009 C57BL/6
B-hCD47 (C) mice 110615 BALB/c
B-hCD47 mice 110010 C57BL/6
B-hCTLA4 mice 110011 C57BL/6
B-hHAVCR2 (TIM3) mice 110018 C57BL/6
B-hLAG3 mice 110025 C57BL/6
B-hOX40L mice 110043 C57BL/6
B-hPD-1 mice 110003 C57BL/6
B-hPD-1 mice Plus 110019 C57BL/6
B-hPD-L1 (CD274) mice 110015 C57BL/6
B-hSIRPα mice 110016 C57BL/6
B-hTIGIT mice 110017 C57BL/6
B-hTNFRSF18 (GITR) mice 110012 C57BL/6
B-hTNFRSF4 (OX40) mice 110014 C57BL/6
B-hTNFRSF9 (4-1BB) mice 110004 C57BL/6

B-NDG-Based Mouse Models

Product Product No Background
B-NDG B2m KO mice plus 110601 B-NDG
B-NDG H2-Ab1 KO mice B-NDG
B-NDG HLA-A2.1 mice 110599 B-NDG
B-NDG hCSF1 mice B-NDG
B-NDG hIL15 mice 110600 B-NDG
B-NDG hIL3 mice B-NDG
B-NDG hIL3/hGM-CSF mice B-NDG
B-NDG hTHPO mice 110590 B-NDG
B-NDG nude mice B-NDG

Double Humanized Immune-checkpoint Mice

Product Product No Background
B-hCTLA4/h4-1BB mice 120539 C57BL/6
B-hCTLA4/hGITR Mice 120962 C57BL/6
B-hCTLA4/hLAG3 mice 120529 C57BL/6
B-hCTLA4/hOX40 mice 120531 C57BL/6
B-hCTLA4/hTIM3 mce 120530 C57BL/6
B-hOX40/h4-1BB mice 120527 C57BL/6
B-hPD-1/h4-1BB mice 120516 C57BL/6
B-hPD-1/hBTLA mice 120517 C57BL/6
B-hPD-1/hCD27 mice 120518 C57BL/6
B-hPD-1/hCD40 mice 120526 C57BL/6
B-hPD-1/hCTLA4 mice 120519 C57BL/6
B-hPD-1/hGITR mice 120528 C57BL/6
B-hPD-1/hLAG3 mice 120520 C57BL/6
B-hPD-1/hOX40 mice 120521 C57BL/6
B-hPD-1/hPD-L1 mice 120522 C57BL/6
B-hPD-1/hTIGIT mice 120523 C57BL/6
B-hPD-1/hTIM3 mice 120524 C57BL/6
B-hPD-L1/h4-1BB mice 120535 C57BL/6
B-hPD-L1/hCD27 mice 120537 C57BL/6
B-hPD-L1/hCD40 mice 120536 C57BL/6
B-hPD-L1/hCTLA4 mice 120538 C57BL/6
B-hPD-L1/hLAG3 mice 120533 C57BL/6
B-hPD-L1/hOX40 mice 120534 C57BL/6
B-hPD-L1/hTIGIT mice 120532 C57BL/6
B-hPD-L1/hTIM3 mice 120540 C57BL/6
B-hSIRPα/hCD47 mice 120525 C57BL/6

Humanized Cytokines Mice

Product Product No Background
B-hIL10 mice 110060 C57BL/6
B-hIL1B mice 110075 C57BL/6
B-hIL2 mice 110085 C57BL/6
B-hIL23A/hIL12B mice 120553 C57BL/6
B-hIL2RA mice 110066 C57BL/6
B-hIL33 mice C57BL/6
B-hIL4 mice 110051 C57BL/6
B-hIL4/IL4RA mice 120551 C57BL/6
B-hIL4RA Mice 110052 C57BL/6
B-hIL6/hIL6R mice 120550 C57BL/6
B-hTNFA mice 110002 C57BL/6

B-NDG Background Mouse Models

Product Product No Background
B-NDG KitW-v mice 110588 B-NDG
B-NDG hSIRPA mice 110003 B-NDG

Humanized Cytokines Mice

Product Product No Background
B-hIL17A Mice 110053 C57BL/6

Other Humanized Mice

Product Product No Background
B-hCCR2 mice 110097 C57BL/6
B-hFcRn mice 110001 C57BL/6
B-hTLR9 mice 110103 C57BL/6

Tetra Humanized Immune-checkpoint Mice

Product Product No Background
B-hPD-1/hPD-L1/hSIRPA/hCD47 mice 140577 C57BL/6

Triple Humanized Immune-checkpoint Mice

Product Product No Background
B-hPD-1/hPD-L1/h4-1BB mice 130569 C57BL/6
B-hPD-1/hPD-L1/hCTLA4 mice 130571 C57BL/6
B-hPD-1/hPD-L1/hLAG3(plus) mice 130572 C57BL/6
B-hPD-1/hPD-L1/hTIM3 mice 130574 C57BL/6
B-hPD-1/hSIRPA/hCD47 mice 130562 C57BL/6
B-hPD-L1/hSIRPA/hCD47 mice 130561 C57BL/6
Product Product No Background
B-HLA-A2.1 Mice 110110 C57BL/6
B-hB7-H3 mice 110028 C57BL/6
B-hC5AR1 Mice 110078 C57BL/6
B-hCCR4 mice 110098 C57BL/6
B-hCD38 mice 110046 C57BL/6
B-hCD39 mice 110792 C57BL/6
B-hCD73 mice 110027 C57BL/6
B-hCD94 Mice 110044 C57BL/6
B-hCSF1R mice 110035 C57BL/6
B-hIL2RB mice 110737 C57BL/6
B-hIL2RG mice 110087 C57BL/6
B-hLAG3 mice plus 110025 C57BL/6
B-hNKG2A mice 110041 C57BL/6
B-hTLR8 Mice 110104 C57BL/6
B-hTNFA/hIL17A mice 120548 C57BL/6
B-hTNFA/hTNFR2 mice 120749 C57BL/6
B-hTNFR2 mice 110032 C57BL/6
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