Mouse Gene Editing

We gene edit several different strains of mice, including C57BL/6, Balb/c, DBA2, and NOD-scid. Models can be generated either by EGE™ or ESC-based gene editing technologies. The turnaround time for CRISPR/Cas9 EGE™-based and ESC-based models is 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. They are also 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. Inbred mice are usually bred to maintain maximum homozygosity. Inbred strains (also called inbred lines) are individuals of a particular species which are 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.

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 B-hCD3ε mice.

Mouse-Protien-Expression-Analysis

Single Humanized Immune-checkpoint Mice

Product Product No Background
B-hBTLA Mice B-CM-018 C57BL/6
B-hCD27 Mice B-EM-024 C57BL/6
B-hCD28 Mice B-CM-019 C57BL/6
B-hCD3e Mice B-EM-026 C57BL/6
B-hCD40 Mice B-CM-020 C57BL/6
B-hCD47 (C) Mice B-CM-022 BALB/c
B-hCD47 Mice B-CM-021 C57BL/6
B-hCTLA4 Mice B-CM-024 C57BL/6
B-hHAVCR2 (TIM3) Mice B-CM-032 C57BL/6
B-hLAG3 Mice B-CM-046 C57BL/6
B-hPD-1 Mice B-CM-001 C57BL/6
B-hPD-1 Mice Plus B-CM-034 C57BL/6
B-hPD-L1 (CD274) Mice B-CM-029 C57BL/6
B-hSIRPα Mice B-CM-030 C57BL/6
B-hTIGIT Mice B-CM-031 C57BL/6
B-hTNFR2 110032 C57BL/6
B-hTNFRSF18 (GITR) Mice B-CM-025 C57BL/6
B-hTNFRSF4 (OX40) Mice B-CM-027 C57BL/6
B-hTNFRSF9 (4-1BB) Mice B-EM-023 C57BL/6

Double Humanized Immune-checkpoint Mice

Product Product No Background
B-hCTLA4/h4-1BB Mice B-DM-024 C57BL/6
B-hCTLA4/hLAG3 Mice B-DM-014 C57BL/6
B-hCTLA4/hOX40 Mice B-DM-016 C57BL/6
B-hCTLA4/hTIM3 Mice B-DM-015 C57BL/6
B-hOX40/h4-1BB Mice B-DM-012 C57BL/6
B-hPD-1/PD-L1 Mice B-DM-007 C57BL/6
B-hPD-1/h4-1BB Mice B-DM-001 C57BL/6
B-hPD-1/hBTLA Mice B-DM-002 C57BL/6
B-hPD-1/hCD27 Mice B-DM-003 C57BL/6
B-hPD-1/hCD40 Mice B-DM-011 C57BL/6
B-hPD-1/hCTLA4 Mice B-DM-004 C57BL/6
B-hPD-1/hGITR Mice B-DM-013 C57BL/6
B-hPD-1/hLAG3 Mice B-DM-005 C57BL/6
B-hPD-1/hOX40 Mice B-DM-006 C57BL/6
B-hPD-1/hTIGIT Mice B-DM-008 C57BL/6
B-hPD-1/hTIM3 Mice B-DM-009 C57BL/6
B-hPD-L1/h4-1BB Mice B-DM-020 C57BL/6
B-hPD-L1/hCD27 Mice B-DM-022 C57BL/6
B-hPD-L1/hCD40 Mice B-DM-021 C57BL/6
B-hPD-L1/hCTLA4 Mice B-DM-023 C57BL/6
B-hPD-L1/hLAG3 Mice B-DM-018 C57BL/6
B-hPD-L1/hOX40 Mice B-DM-019 C57BL/6
B-hPD-L1/hTIGIT Mice B-DM-017 C57BL/6
B-hPD-L1/hTIM3 Mice B-DM-026 C57BL/6
B-hSIRPα/hCD47 Mice B-DM-010 C57BL/6

Triple Humanized Immune-checkpoint Mice

Product Product No Background
B-hPD-1/hPD-L1/hCTLA4 130571 C57BL/6
B-hPD-1/hPD-L1/hTIM3 130574 C57BL/6
B-hPD-1/hSIRPA/hCD47 Mice B-TM-002 C57BL/6
B-hPD-L1/hSIRPA/hCD47 Mice B-TM-001 C57BL/6

Cre Mouse Models List

Product Product No Application Fields
B-ALB-CreERT2 Mice B-CM-014 Alb promoter-driven inducible Cre mouse model (liver)
B-CAG-tdTomato cKI Mice B-CM-066 Cre-induced tdTomato expression, for Cre expression profile detection
B-CD150-iCreERT2 Mice B-CM-175 CD150 promoter-driven inducible Cre mouse model (Thymic, T cells and macrophages)
B-Col10a1-iCre Mice B-EM-001 Col10a1 promoter-driven Cre mouse model (mature hypertrophic chondrocyte)
B-Col2a1-iCre Mice B-EM-002 Col2 promoter-driven Cre mouse model (type II collencyte)
B-Dmp1-iCre Mice B-EM-003 Dmp1 promoter-driven Cre mouse model (skeleton)
B-Grpr-iCre Mice B-EM-004 Grpr promoter-driven Cre mouse model (digestive system, nervous system)
B-Ins1-iCreERT2 Mice B-EM-005 Ins1 promoter-driven inducible Cre mouse model (pancreas isletbeta cell)
B-Ncr1 (Nkp46)-iCre Mice B-EM-006 Ncr1 promoter-driven Cre mouse model (NK cell)
B-Oxr1-iCreERT2 Mice B-CM-003 Oxr1 promoter-driven inducible Cre mouse model (frontal lobe adult, cerebellum adult and 24 other tissues)
B-Pdgfrb-iCre Mice B-CM-004 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 B-EM-007 Prrx1 promoter-driven Cre mouse model (mesenchyme)
B-Sp7 (Osx)-iCre Mice B-EM-008 Sp7 promoter-driven Cre mouse model (skeleton)
B-Syn1-iCre Mice B-CM-005 Syn1promoter-driven inducible Cre mouse model (brain)
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 B-EM-010 UCP1 promoter-driven Cre mouse model (mitochondria, brown fat)
B-Zeb1-iCreERT2 Mice B-EM-011 Zeb1 promoter-driven inducible Cre mouse model (tunica media of artery and myocardial cell)

Other Genetically Modified Mouse Models

Product Product No Application Fields
B-Apoe-KO Mice B-EM-021 Hyperlipidemia, atherosclerosis mice model (Cardiovascular system)
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-hIL17A Mice B-EM-038
B-hIL4/IL4R Mice B-DM-025
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
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