B-hCD16A/hCD20 mice

C57BL/6N-Fcgr4tm2(FCGR3A)Bcgen Ms4a1tm2(MS4A1)Bcgen/Bcgen • 113998

B-hCD16A/hCD20 mice

Product nameB-hCD16A/hCD20 mice
Catalog number113998
Strain nameC57BL/6N-Fcgr4tm2(FCGR3A)Bcgen Ms4a1tm2(MS4A1)Bcgen/Bcgen
Strain backgroundC57BL/6N
AliasesFCG3, CD16A, FCGR3, IGFR3, IMD20, CD16-II, FCRIIIA, FcGRIIIA; B1, S7, Bp35, CD20, FMC7, CVID5, LEU-16

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  • Targeting strategy
  • Phenotypic analysis

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    发表文章

      Targeting strategy

      CD16A

      • The exons 1-5 of mouse Fcgr4 gene that encode signal peptide, extracellular domain, transmembrane domain, and cytoplasmic region are replaced by human FCGR3A(F158V) exons 1-6 in B-hCD16A/hCD20 mice.
      • The promoter and 5’UTR region of the mouse gene are replaced by human counterparts. The human FCGR3A expression is driven by human FCGR3A promoter, while mouse Fcgr4 gene transcription and translation will be disrupted.

      CD20

      • The exons 2-7 of mouse Cd20 gene that encode the whole molecule (ATG to STOP codon) are replaced by human counterparts in B-hCD16A/hCD20 mice.
      • The promoter, 5’UTR, and 3’UTR region of the mouse gene are retained. The human CD20 expression is driven by endogenous mouse Cd20 promoter, while mouse Cd20 gene transcription and translation will be disrupted.
      CD16A Protein Expression Analysis
      • Mouse CD16.2 was detected in wild-type  C57BL/6JNifdc mice, but not in B-hCD16A/hCD20 mice.
      • Human CD16A was exclusively detected in homozygous B-hCD16A/hCD20 mice.

      Mouse and human CD16A expression analysis in splenocytes. Splenocytes were collected from wild-type (WT) C57BL/6JNifdc mice, homozygous (HO) B-hCD16A/hCD20 mice. CD16A expression on various types of immune cells was analyzed by flow cytometry using species-specific anti-mouse CD16.2 antibody (Biolegend, 149503) and anti-human CD16A antibody (BioLegend, 360706).

      • Mouse CD16.2 was detected in wild-type  C57BL/6JNifdc mice, but not in B-hCD16A/hCD20 mice.
      • Human CD16A was exclusively detected in homozygous B-hCD16A/hCD20 mice.

      Mouse and human CD16A expression analysis in splenocytes. Splenocytes were collected from wild-type (WT) C57BL/6JNifdc mice, homozygous (HO) B-hCD16A/hCD20 mice. CD16A expression on various types of immune cells was analyzed by flow cytometry using species-specific anti-mouse CD16.2 antibody (Biolegend, 149503) and anti-human CD16A antibody (BioLegend, 360706).

      • Mouse CD16.2 was detected in wild-type  C57BL/6JNifdc mice, but not in B-hCD16A/hCD20 mice.
      • Human CD16A was exclusively detected in homozygous B-hCD16A/hCD20 mice.

      Mouse and human CD16A expression analysis in splenocytes. Blood cells were collected from wild-type (WT) C57BL/6JNifdc mice, homozygous (HO) B-hCD16A/hCD20 mice. CD16A expression on various types of immune cells was analyzed by flow cytometry using species-specific anti-mouse CD16.2 antibody (Biolegend, 149503) and anti-human CD16A antibody (BioLegend, 360706).

      • Mouse CD16.2 was detected in wild-type  C57BL/6JNifdc mice, but not in B-hCD16A/hCD20 mice.
      • Human CD16A was exclusively detected in homozygous B-hCD16A/hCD20 mice.

      Mouse and human CD16A expression analysis in splenocytes. Blood cells were collected from wild-type (WT) C57BL/6JNifdc mice, homozygous (HO) B-hCD16A/hCD20 mice. CD16A expression on various types of immune cells was analyzed by flow cytometry using species-specific anti-mouse CD16.2 antibody (Biolegend, 149503) and anti-human CD16A antibody (BioLegend, 360706).

      CD20 Protein Expression Analysis
      • Mouse CD20 was detected in wild-type C57BL/6JNifdc mice, but not in B-hCD16A/hCD20 mice.
      • Human CD20 was exclusively detected in homozygous B-hCD16A/hCD20 mice.

      Mouse and human CD20 expression analysis in splenocytes and blood cells. Splenocytes and blood cells were collected from wild-type (WT) C57BL/6JNifdc mice, homozygous (HO) B-hCD16A/hCD20 mice. CD20 expression was analyzed by flow cytometry using species-specific anti-mouse CD20 antibody (Biolegend, 152107) and anti-human CD20 antibody (BioLegend, 375504).

      Analysis of Leukocyte Subpopulations

      Analysis of leukocyte subpopulations by flow cytometry in immune organs and blood. Splenocytes, peripheral blood, and lymph nodes were isolated from female C57BL/6JNifdc mice and B-hCD16A/hCD20 mice (female, 8-week-old, n = 3). Single live cells were gated on the CD45⁺ population and analyzed by flow cytometry as indicated. Values are expressed as mean ± SEM.

      Analysis of T Cell Subpopulations
      • The frequencies of CD4⁺ T cells, CD8⁺ T cells, and Tregs in homozygous B-hCD16A/hCD20 mice were comparable to those in C57BL/6JNifdc mice.
      • Humanization of CD16A, and CD20 does not affect normal T cell development, differentiation, or splenic distribution.

      Analysis of T-cell subpopulations by flow cytometry in immune organs and blood. Splenocytes, peripheral blood, and lymph nodes were isolated from female C57BL/6JNifdc mice and B-hCD16A/hCD20 mice (female, 8-week-old, n = 3). Single live cells were gated on the CD3⁺ T-cell population and analyzed by flow cytometry as indicated. Values are expressed as mean ± SEM.

      * When publishing results obtained using this animal model, please acknowledge the source as follows: The animal model [B-hCD16A/hCD20 mice] (Cat# 113998) was purchased from Biocytogen.
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