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Using mouse models to study function of transcriptional factors in T cell development.

Li P, Xiao Y, Liu Z, Liu P - Cell Regen (Lond) (2012)

Bottom Line: With the advances of genetic engineering and conditional knockout (CKO) mice, we now understand hematopoiesis is a dynamic stepwise process starting from hematopoietic stem cells (HSCs) which are responsible for replenishing all blood cells.Transcriptional factors play important role in hematopoiesis.Finally, we focused on the key transcriptional factor Bcl11b and its function in regulating T cell specification and commitment.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Regenerative Biology, Guangzchou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China ; Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou, China.

ABSTRACT
Laboratory mice have widely been used as tools for basic biological research and models for studying human diseases. With the advances of genetic engineering and conditional knockout (CKO) mice, we now understand hematopoiesis is a dynamic stepwise process starting from hematopoietic stem cells (HSCs) which are responsible for replenishing all blood cells. Transcriptional factors play important role in hematopoiesis. In this review we compile several studies on using genetic modified mice and humanized mice to study function of transcriptional factors in lymphopoiesis, including T lymphocyte and Natural killer (NK) cell development. Finally, we focused on the key transcriptional factor Bcl11b and its function in regulating T cell specification and commitment.

No MeSH data available.


Related in: MedlinePlus

General procedure for generation of a conditional knockout mouse strain by gene targeting strategies. (A) Generation of a targeting vector containing critical exon of targeted genes (red rectangle), two loxP site (green triangle), a positive (+) selection cassette and sequences of homology with the target locus (blue line). (B) The vector is linearized and electroporated into ES cells. (C) Correct tranformants are selected for in the presence of a selectant (eg. G418 if a neomycin resistance cassette is present in the targeting vector). (D) Correctly targeted ES cell clones are then identified and genetically characterized using long range PCR or Southern blot analysis. (E) The selected ES cell clones are then microinjected into 3.5 dpc blastocysts and transplanted into the uteri of pseudopregnant females. (F) Chimeras obtained from the microinjections are mated with wild-type mice to establish germ-line transmission of the modified allele. (G) Progeny derived from the chimeras are characterized using long range PCR or Southern blot analysis, and a mutant mouse line that carries the desired targeted allele is established.
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Fig1: General procedure for generation of a conditional knockout mouse strain by gene targeting strategies. (A) Generation of a targeting vector containing critical exon of targeted genes (red rectangle), two loxP site (green triangle), a positive (+) selection cassette and sequences of homology with the target locus (blue line). (B) The vector is linearized and electroporated into ES cells. (C) Correct tranformants are selected for in the presence of a selectant (eg. G418 if a neomycin resistance cassette is present in the targeting vector). (D) Correctly targeted ES cell clones are then identified and genetically characterized using long range PCR or Southern blot analysis. (E) The selected ES cell clones are then microinjected into 3.5 dpc blastocysts and transplanted into the uteri of pseudopregnant females. (F) Chimeras obtained from the microinjections are mated with wild-type mice to establish germ-line transmission of the modified allele. (G) Progeny derived from the chimeras are characterized using long range PCR or Southern blot analysis, and a mutant mouse line that carries the desired targeted allele is established.

Mentions: In 1981, pluripotent mouse embryonic stem cells (ES) were isolated from the inner cell mass of 3.5 days post-coitum (dpc) wild type mouse embryos [19, 20]. Later, it was demonstrated that these ES cells were able to contribute to the germ line in chimera mice derived from ES cells, even after genome modification by retrovirus [21, 22]. Precise manipulation of the mouse genome was achieved by demonstrating that homologous recombination works efficiently in mouse ES cells [23, 24]. Thus a combination of mouse ES cell manipulation and homologous recombination technologies gave birth to ‘gene targeting’ which has revolutionized mouse genetics. With birth of genetically engineered ‘knockout’ mice, target mutations have been introduced to many loci to study gene functions in the immunity. In 1990, Smithies’ and Jaenisch’s group independently generated β2-microglobulin knockout mice in which expression of major histocompatibility complex (MHC) class I molecules was abolished. The mutant mice developed normally but had severely reduced number of CD8+ cytotoxic T lymphocytes (CTLs), indicating that MHC class I molecules are required for the selection of MHC class-I-restricted CD8+ T cells and for antigen recognition by these cells, but not necessary for T cell development in general [25, 26]. It is now feasible to knockout every single gene in the mouse and observe phenotype in this knockout mice (Figure 1) [27].Figure 1


Using mouse models to study function of transcriptional factors in T cell development.

Li P, Xiao Y, Liu Z, Liu P - Cell Regen (Lond) (2012)

General procedure for generation of a conditional knockout mouse strain by gene targeting strategies. (A) Generation of a targeting vector containing critical exon of targeted genes (red rectangle), two loxP site (green triangle), a positive (+) selection cassette and sequences of homology with the target locus (blue line). (B) The vector is linearized and electroporated into ES cells. (C) Correct tranformants are selected for in the presence of a selectant (eg. G418 if a neomycin resistance cassette is present in the targeting vector). (D) Correctly targeted ES cell clones are then identified and genetically characterized using long range PCR or Southern blot analysis. (E) The selected ES cell clones are then microinjected into 3.5 dpc blastocysts and transplanted into the uteri of pseudopregnant females. (F) Chimeras obtained from the microinjections are mated with wild-type mice to establish germ-line transmission of the modified allele. (G) Progeny derived from the chimeras are characterized using long range PCR or Southern blot analysis, and a mutant mouse line that carries the desired targeted allele is established.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230505&req=5

Fig1: General procedure for generation of a conditional knockout mouse strain by gene targeting strategies. (A) Generation of a targeting vector containing critical exon of targeted genes (red rectangle), two loxP site (green triangle), a positive (+) selection cassette and sequences of homology with the target locus (blue line). (B) The vector is linearized and electroporated into ES cells. (C) Correct tranformants are selected for in the presence of a selectant (eg. G418 if a neomycin resistance cassette is present in the targeting vector). (D) Correctly targeted ES cell clones are then identified and genetically characterized using long range PCR or Southern blot analysis. (E) The selected ES cell clones are then microinjected into 3.5 dpc blastocysts and transplanted into the uteri of pseudopregnant females. (F) Chimeras obtained from the microinjections are mated with wild-type mice to establish germ-line transmission of the modified allele. (G) Progeny derived from the chimeras are characterized using long range PCR or Southern blot analysis, and a mutant mouse line that carries the desired targeted allele is established.
Mentions: In 1981, pluripotent mouse embryonic stem cells (ES) were isolated from the inner cell mass of 3.5 days post-coitum (dpc) wild type mouse embryos [19, 20]. Later, it was demonstrated that these ES cells were able to contribute to the germ line in chimera mice derived from ES cells, even after genome modification by retrovirus [21, 22]. Precise manipulation of the mouse genome was achieved by demonstrating that homologous recombination works efficiently in mouse ES cells [23, 24]. Thus a combination of mouse ES cell manipulation and homologous recombination technologies gave birth to ‘gene targeting’ which has revolutionized mouse genetics. With birth of genetically engineered ‘knockout’ mice, target mutations have been introduced to many loci to study gene functions in the immunity. In 1990, Smithies’ and Jaenisch’s group independently generated β2-microglobulin knockout mice in which expression of major histocompatibility complex (MHC) class I molecules was abolished. The mutant mice developed normally but had severely reduced number of CD8+ cytotoxic T lymphocytes (CTLs), indicating that MHC class I molecules are required for the selection of MHC class-I-restricted CD8+ T cells and for antigen recognition by these cells, but not necessary for T cell development in general [25, 26]. It is now feasible to knockout every single gene in the mouse and observe phenotype in this knockout mice (Figure 1) [27].Figure 1

Bottom Line: With the advances of genetic engineering and conditional knockout (CKO) mice, we now understand hematopoiesis is a dynamic stepwise process starting from hematopoietic stem cells (HSCs) which are responsible for replenishing all blood cells.Transcriptional factors play important role in hematopoiesis.Finally, we focused on the key transcriptional factor Bcl11b and its function in regulating T cell specification and commitment.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Regenerative Biology, Guangzchou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China ; Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou, China.

ABSTRACT
Laboratory mice have widely been used as tools for basic biological research and models for studying human diseases. With the advances of genetic engineering and conditional knockout (CKO) mice, we now understand hematopoiesis is a dynamic stepwise process starting from hematopoietic stem cells (HSCs) which are responsible for replenishing all blood cells. Transcriptional factors play important role in hematopoiesis. In this review we compile several studies on using genetic modified mice and humanized mice to study function of transcriptional factors in lymphopoiesis, including T lymphocyte and Natural killer (NK) cell development. Finally, we focused on the key transcriptional factor Bcl11b and its function in regulating T cell specification and commitment.

No MeSH data available.


Related in: MedlinePlus