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 Immune-checkpoint Mice

Product Product No Background
B-hPD-1/hPD-L1/h4-1BB mice 130569 C57BL/6
B-hPD-1/hPD-L1/hGITR mice 131108 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

Double Humanized Immune-checkpoint Mice

Product Product No Background
B-h4-1BB/h4-1BBL mice 120541 C57BL/6
B-hCD94/hNKG2A mice 121077 C57BL/6
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-hIL12A/hIL12B mice 120552 C57BL/6
B-hOX40/h4-1BB mice 120527 C57BL/6
B-hPD-1/h4-1BB mice 120516 C57BL/6
B-hPD-1/hB7-H3 mice 121152 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

Single Humanized Immune-checkpoint Mice

Product Product No Background
B-dPD-1 mice 110022 C57BL/6
B-h4-1BB mice 110004 C57BL/6
B-hB7-H3 mice 110028 C57BL/6
B-hBTLA mice 110005 C57BL/6
B-hCCR8 mice 110096 C57BL/6
B-hCD24 mice 110767 C57BL/6
B-hCD27 mice 110006 C57BL/6
B-hCD28 mice 110007 C57BL/6
B-hCD33 mice 110823 C57BL/6
B-hCD38 mice 110046 C57BL/6
B-hCD3E mice 110008 C57BL/6
B-hCD40 mice 110009 C57BL/6
B-hCD44 mice 110961 C57BL/6
B-hCD47 mice 110010 C57BL/6
B-hCD47 mice(C) 110615 BALB/c
B-hCD52 mice 110817 C57BL/6
B-hCD73 mice plus 110047 C57BL/6
B-hCD79B mice 111096 C57BL/6
B-hCTLA4 mice 110011 C57BL/6
B-hGARP mice 110102 C57BL/6
B-hGITR mice 110012 C57BL/6
B-hGLP1R mice 110106 C57BL/6
B-hLAG3 mice 110013 C57BL/6
B-hOX40 mice 110014 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 mice 110015 C57BL/6
B-hSIGLEC10 mice 110843 C57BL/6
B-hSIGLEC15 mice 110121 C57BL/6
B-hSIRPA mice 110016 C57BL/6
B-hTIGIT mice 110017 C57BL/6
B-hTIM3 mice 110018 C57BL/6
B-hTIM4 mice 110837 C57BL/6
B-hVSIR mice 110049 C57BL/6

B-NDG-Based Mouse Models

Product Product No Background
B-NDG B2m KO mice plus 110601 B-NDG
B-NDG HLA-A2.1 mice 110599 B-NDG
B-NDG hIL15 mice 110600 B-NDG
B-NDG hSIRPA/hCD47 mice 110603 B-NDG
B-NDG hTHPO mice 110590 B-NDG

B-NDG Background Mouse Models

Product Product No Background
B-NDG KitW-v mice 110588 B-NDG
B-NDG hCSF1/hTHPO mice 120609 B-NDG
B-NDG hSIRPA mice 110604 B-NDG

Triple Humanized

Product Product No Background
B-hPD-1/hLAG3/hTIM3 mice 130999 C57BL/6
B-hPD-1/hPD-L1/hCD40 mice 131083 C57BL/6
B-hPD-1/hPD-L1/hCD73 plus mice 131036 C57BL/6
B-hPD-1/hPD-L1/hCTLA4 mice 130571 C57BL/6
B-hPD-1/hPD-L1/hTIGIT mice 130573 C57BL/6
B-hPD-1/hPD-L1/hTNFR2 mice 130849 C57BL/6

Other Genetically Modified Mouse Models

Product Product No Application Fields
B-Apoe KO mice 110168 Hyperlipidemia, atherosclerosis mice model (Cardiovascular system)
B-F8 KO mice 110169 Hemophilia mice model
B-F9 KO mice 110170 Hemophilia mice model
B-Il17a-EGFP mice 110161 IL17 expression and functional test (gene reporter mice)
B-PD-1-EGFP KI mice 110166 PD-1, PD1
B-Tgfb1 cKO mice 110164 Study on transforming growth factor
B-ob/ob mice 110172 Diabetes mice model
B-p53 KO mice 110167 Tumorigenesis drug test

Humanized Cytokines Mice

Product Product No Background
B-hIL10 mice 110060 C57BL/6
B-hIL1B mice 110075 C57BL/6
B-hIL1RAcP mice 110072 C57BL/6
B-hIL2 mice 110085 C57BL/6
B-hIL23A/hIL12B mice 120553 C57BL/6
B-hIL2RA mice 110066 C57BL/6
B-hIL33 mice 110055 C57BL/6
B-hIL4 mice 110051 C57BL/6
B-hIL4/hIL4RA mice 120551 C57BL/6
B-hIL4RA Mice 110052 C57BL/6
B-hIL6/hIL6R mice 120557 C57BL/6
B-hTNFA mice 110002 C57BL/6

Cre Mouse Models List

Product Product No Application Fields
B-CAG-tdTomato cKI mice 110148 Cre-induced tdTomato expression, for Cre expression profile detection
B-Gfap-iCre mice 110144 Gfappromoter-driven Cre mouse model. (Astroglia)
B-Grpr-iCre Mice 110125 Grpr promoter-driven Cre mouse model (digestive system, nervous system)
B-Nkp46-iCre mice 110127 Ncr1 promoter-driven Cre mouse model (NK cell)
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-iCre mice 110131 Sp7 promoter-driven Cre mouse model (skeleton)
B-Tcf21-iCreERT2 mice 110141 Tcf21 promoter-driven inducible Cre mouse model (bladder, lung)
B-Ucp1-iCre Mice 110134 UCP1 promoter-driven Cre mouse model (mitochondria, brown fat)

Humanized Cytokines Mice

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

Tetra Humanized Immune-checkpoint Mice

Product Product No Background
B-hPD-1/hPD-L1/hSIRPA/hCD47 mice 140577 C57BL/6
Product Product No Background
B-HLA-A2.1 Mice 110110 C57BL/6
B-hCCR5 mice 110099 C57BL/6
B-hCD226 mice 110869 C57BL/6
B-hCD39 mice 110783 C57BL/6
B-hCD3EDG mice 110039 C57BL/6
B-hCD5 mice 111125 C57BL/6
B-hCD73 mice 110027 C57BL/6
B-hCD94 Mice 110044 C57BL/6
B-hCSF1/hCSF1R mice 121109 C57BL/6
B-hCSF1R mice 110035 C57BL/6
B-hCSF2RB mice 110735 C57BL/6
B-hIL13 mice 110058 C57BL/6
B-hIL15RA mice 110112 C57BL/6
B-hIL17A/hIL17F mice 120554 C57BL/6
B-hIL1A mice 110073 C57BL/6
B-hIL21R mice 110766 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-hPD-1/hPD-L1/hIL2RA mice 131078 C57BL/6
B-hPD-1/hPD-L1/hOX40 mice 311112 C57BL/6
B-hPSGL-1 mice 111470 C57BL/6
B-hRAGE mice 111107 C57BL/6
B-hRANKL mice 110772 C57BL/6
B-hTLR8 Mice 110104 C57BL/6
B-hTNFA/hIL17A mice 120548 C57BL/6
B-hTNFR2 mice 110032 C57BL/6
B-hTNFR2/hTNFR1 mice 120802 C57BL/6
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