B-Cd19-EGFP-DTR-luciferase mice

Basic Information

Strain name
C57BL/6JNifdc-Cd19tm1(EGFP-DTR-luc)Bcgen/Bcgen
Common name
B-Cd19-EGFP-DTR-luciferase mice
Background
C57BL/6JNifdc
Catalog number
112817
Aliases
B4, CVID3
NCBI gene ID

Targeting strategy

Gene targeting strategy for B-Cd19-EGFP-DTR-luciferase mice. 

A construct composed of the cDNA for enhanced GFP (EGFP), the human DTR, and luciferase was inserted before the stop codon of the Cd19 gene in B-Cd19-EGFP-DTR-luciferase mice, allowing for EGFP, DTR and luciferase expression and de novo Cd19 expression. The coding sequences were separated by self-cleaving 2A peptide sequences.

EGFP expression analysis in leukocytes cell subpopulation

Frequencies of EGFP in leukocytes cell subpopulation from spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old). Flow cytometry analysis of the splenocytes, blood and lymph node was performed to assess the frequencies of EGFP+ cells in leukocytes cell subpopulation from spleen(A), blood (B) and lymph node(C). EGFP+ cells was expressed at a high level in B cells from spleen, blood and lymph node in heterozygous mice. And there was no leaky expression of EGFP in other cell subpopulation.

CD19+ cells depletion analysis

Frequencies of CD19+; EGFP cells in spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the splenocytes, blood and lymph node was performed to assess the frequencies of CD19+; EGFP+ cells in total CD45+ cells. The signal for CD19+;EGFP+ cells in spleen(A), in blood (B) and lymph node(C) were significantly decreased in heterozygous mice injected with DT.

CD19+ cells depletion analysis in spleen

Frequencies of CD19+ cells and EGFP+ cells in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequencies of CD19+ cells and EGFP+ cells in total CD45+ cells. The signal for CD19+ cells(A) and EGFP+ cells(B) were decreased in heterozygous mice injected with DT. C. Percentages of CD19+ cells and EGFP+ cells in total CD45+ populations. The frequencies of CD19+ cells and EGFP+ cells were significantly decreased in heterozygous mice after DT injection.

CD19+ cells depletion analysis in blood

Frequencies of CD19+ cells and EGFP+ cells in blood by flow cytometry. Blood were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the blood was performed to assess the frequencies of CD19+ cells and EGFP+ cells in total CD45+ cells. The signal for CD19+ cells(A) and EGFP+ cells(B) were decreased in heterozygous mice injected with DT. C. Percentages of CD19+ cells and EGFP+ cells in total CD45+ populations. The frequencies of CD19+ cells and EGFP+ cells were significantly decreased in heterozygous mice after DT injection.

CD19+ cells depletion analysis in lymph node

Frequencies of CD19+ cells and EGFP+ cells in lymph node by flow cytometry. Lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the lymph node was performed to assess the frequencies of CD19+ cells and EGFP+ cells in total CD45+ cells. The signal for CD19+ cells(A) and EGFP+ cells(B) were decreased in heterozygous mice injected with DT. C. Percentages of CD19+ cells and EGFP+ cells in total CD45+ populations. The frequencies of CD19+ cells and EGFP+ cells were significantly decreased in heterozygous mice after DT injection.

B cells depletion analysis

Frequencies of B cells in spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the splenocytes, blood and lymph node was performed to assess the frequencies of B cells in total CD45+ cells. The signal for B cells in spleen(A), blood (B) and lymph node(C) were significantly decreased in heterozygous mice injected with DT.

Frequencies of EGFP in B cells from spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the splenocytes, blood and lymph node was performed to assess the frequencies of EGFP cells in total B cells. The signal for EGFP in B cells in spleen(A), blood (B) and lymph node(C) were significantly decreased in heterozygous mice injected with DT.

Analysis of B cells depletion in spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. The percentages of B cells in total CD45 cells from spleen, blood and lymph node(A) and EGFP+ cells in B cells from spleen, blood and lymph node(B) were significantly decreased in heterozygous mice injected with DT.

Frequency of leukocyte subpopulations in spleen

Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice and B-Cd19-EGFP-DTR-luc mice(n=3, 6-7-week-old). Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in heterozygous B-Cd19-EGFP-DTR-luc mice were similar to those in C57BL/6 mice. The frequency of leukocyte subpopulations in blood and lymph node of B-Cd19-EGFP-DTR-luc mice were also comparable to wild-type C57BL/6 mice (Data not shown). Values are expressed as mean ± SEM.

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