Overview

Also Known As:Jedi (Just EGFP Death Inducing), Jedi

Jedi (Just EGFP Death Inducing) mice are a GFP-specific TCR mouse line that produces T cells that recognize the immunodominant epitope of enhanced GFP (GFP_200-208) presented on H-2K^d; enabling GFP to be used as a model antigen. Activation of Jedi CD8⁺ T cells rapidly and stably eliminates GFP⁺ cell populations. Jedi are the first antigen-specific TCR mice with T cells recognizing a florescent protein. The ability for direct fluorescent visualization of the target antigen is a novel and invaluable aspect of the Jedi technology.

Donating Investigator

Brian D Brown, Icahn School of Medicine at Mount Sinai , Mount Sinai School of Medicine

Genetic overview

Genetic Background	Generation
	?+F4 (2020-07-28 00:00:00)

Tcrb^Ln1Bdb

Allele Type	Gene Symbol	Gene Name
Other (Not Specified)	Tcrb	T cell receptor beta chain

H2^d

Allele Type	Gene Symbol	Gene Name
Spontaneous	H2	histocompatibility-2, MHC

Ptprc^a

Allele Type	Gene Symbol	Gene Name
Not Applicable	Ptprc	protein tyrosine phosphatase, receptor type, C

Tcra^Ln1Bdb

Allele Type	Gene Symbol	Gene Name
Other (Not Specified)	Tcra	T cell receptor alpha chain

View Genetics

Research Applications

Research Tools
Immunology, Inflammation and Autoimmunity Research

View All Research Applications

Base Price

Starting at:

$255.00 Domestic price for female 4-week

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Details

Detailed Description

The GFP-specific TCR mouse line Jedi (Just EGFP Death Inducing) produces T cells that recognize the immunodominant epitope of enhanced GFP (GFP_200-208) presented on H-2K^d. In every Jedi mouse, at least 50% of CD8⁺ T cells are specific for the GFP_200-208-H-2K^d pentamer and their phenotype is naïve (CD44^- CD62L⁺).
Specificity is high: Jedi CD8⁺ T-cells target and destroy GFP⁺ cells, but not YFP⁺ cells (despite that YFP has >95% homology to GFP but differs by one amino acid in the immunodominant epitope). Similarly, when co-cultured with dendritic cells expressing GFP or YFP, the Jedi CD8⁺ T cells proliferate in the presence of GFP but not YFP.

Jedi CD8⁺ T cells rapidly and stably eliminate GFP⁺ cell populations. Whether GFP mice are designed to express GFP on the cell surface or in the cell interior (cytoplasm, or bound to the inner side of membranes, etc.), all GFP expressing cells (with normal MHC class I function) present some GFP epitopes on their MHC class I molecules - that random occurrence of "self GFP_200-208 epitope" presentation on those cells is enough for Jedi T cells to identify them. Some examples are listed below.
When Jedi CD8⁺ T cells are injected into mice expressing GFP in pancreatic β-cells (MIP-EGFP; see Stock No. 006864), subsequent vaccination with GFP activates the transferred cells - resulting in rapid β-cell destruction and hyperglycemia. This is a model of inducible type I diabetes that also allows direct visualization of the islet-antigen. [Agudo et al. 2015 Nat Biotechnol 33:1287]
When Jedi CD8⁺ T cells are injected into mice engineered to express GFP in microglia (CX3CR1-EGFP; see Stock No. 005582) but RFP in peripheral hematopoietic cells, the resulting elimination of the GFP⁺ microglia occurred rapidly both with or without GFP vaccination/activation. This is a model for studying T cell trafficking and cytotoxic lymphocyte clearance. [Agudo et al. 2015 Nat Biotechnol 33:1287]
Jedi CD8⁺ T cells are able to stably deplete a population of renewing cells. For example, when injected into mice expressing GFP in Foxp3⁺ regulatory T cells (Foxp3-EGFP; similar to Stock Nos. 006772 or 018628), subsequent vaccination with GFP activates the transferred cells - resulting in rapid and stable depletion of Foxp3⁺GFP⁺ Tregs. Specifically, when the host is a female heterozygous for the X-linked Foxp3-EGFP allele, only ~half of the Foxp3⁺ Tregs are GFP⁺ due to random X inactivation. While those cells are stably depleted, the Foxp3⁺GFP^- cells remained and proliferated to compensate for the loss of the GFP⁺ Tregs. When the host is a male hemizygous for the X-linked Foxp3-EGFP allele, complete depletion of Foxp3⁺ Tregs resulted in immune dysregulation and illness. [Agudo et al. 2015 Nat Biotechnol 33:1287]
Jedi CD8⁺ T cells are able to eliminate rare cell populations in a non-lymphoid, non-inflamed organ. For example, when injected into mice expressing GFP in an extremely rare population of pacemaker heart cells (Hcn4-EGFP), subsequent vaccination with GFP activates the transferred cells - resulting in rapid and stable depletion of Hcn4⁺ cells. Unique from other depletion technologies, this allowed assessment of target cell depletion at single cell resolution via flow cytometry. [Agudo et al. 2015 Nat Biotechnol 33:1287]

When homozygous for the CD45.1 allele (Ptprc^a) on chromosome 1, heterozygous for the two Jedi TCR alleles (Tcrb^Ln1Bdb on chromosome 6 and Tcra^Ln1Bdb on chromosome 14) and homozygous for the H-2K^d allele (H2^d) on chromosome 17, the Jedi mice are viable and fertile. The donating investigator reports that when they breed heterozygous Jedi mice together they obtain the expected number of litters, but their genotyping protocol cannot properly distinguish heterozygous from homozygous to date (August 2018). By November 2019, The Jackson Laboratory has genotyping assays in place for the two Jedi TCR alleles to distinguish heterozygous from homozygous. Additionally, the donating investigator suggests that mice may be immunocompromised when homozygous for the two Jedi TCR alleles.

Development

Jedi (Just EGFP Death Inducing) mice harbor four alleles that are not linked and are expected to segregate independently: the CD45.1 allele (Ptprc^a) on chromosome 1, the two Jedi TCR alleles (Tcrb^Ln1Bdb on chromosome 6 and Tcra^Ln1Bdb on chromosome 14) and the H-2K^d allele (H2^d) on chromosome 17. Specific details are below.

The two Jedi TCR alleles were created by Dr. Brian D. Brown (Icahn School of Medicine at Mount Sinai) using a somatic cell nuclear transfer (SCNT) approach. First, a lentivirus encoding GFP was injected into (BALB/cJ x C57BL/6J)F1 mice (CB6F1; H-2K^b/d). After 2 weeks, GFP_200-208-H-2K^d tetramer⁺ CD8⁺ T cells were isolated. The donating investigator reports the identity of the T cell receptor Vα and Vβ chains was found to be trav7-4, traj30 and trbv2, trbj1.6 (Vα1-J30 and Vβ4-D1-J1.6-C1). The T cells nucleus was injected into an enucleated oocyte, and the resulting cell was used to generate embryonic stem (ES) cells. ES cells were injected into recipient blastocysts, and blastocysts implanted into psuedopregnant females. The F1 mice were then bred for several generations to the C5-functional B10D2 mice (originally from Stock No. 000463; B10.D2-Hc¹ H2^d H2-T18^c/nSnJ) and selected to retain the H2^d allele (H-2K^d) on chromosome 17. Next, Jedi mice were bred to B6.SJL-Ptprc^a Pepc^b/BoyJ mice (originally from Stock No. 002014) to obtain the Ptprc^a allele (CD45.1) on chromosome 1. The Jedi mice were then bred approximately four more generations to the C5-functional B10D2 mice (originally from Stock No. 000463) and selected to retain the H-2K^d and CD45.1 alleles - resulting in the Jedi H-2K^d CD45.1 colony (on a predominantly B10.D2 genetic background). In 2018, both female and male mice that were homozygous for CD45.1, heterozygous or homozygous for the Jedi TCR alleles and homozygous for H-2K^d were sent to The Jackson Laboratory Repository. Upon arrival, our live colony was generated using those females and males.

Control Suggestions

Approximate Controls

Additional Information

Considerations for Choosing Controls

Selected References

Agudo J; Ruzo A; Park ES; Sweeney R; Kana V; Wu M; Zhao Y; Egli D; Merad M; Brown BD. 2015. GFP-specific CD8 T cells enable targeted cell depletion and visualization of T-cell interactions. Nat Biotechnol 33(12):1287-1292PubMed: 26524661 MGI: J:248646

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Genetics

Tcrb^Ln1Bdb

Allele Symbol: Tcrb^Ln1Bdb

Allele Name	recombination line 1, Brian Brown
Allele Type	Other (Not Specified)
Allele Synonym(s)
Gene Symbol and Name	Tcrb, T cell receptor beta chain
Gene Synonym(s)
Strain of Origin	(BALB/cJ x C57BL/6J)F1
Chromosome	6
Molecular Note	This allele is generated using somatic cell nuclear transfer (SCNT). First, a lentivirus encoding GFP was injected into (BALB/cJ x C57BL/6J)F1 mice (CB6F1; H-2K). After 2 weeks, GFP-200-208- H-2K tetramer<+> CD8<+> T cells were isolated. The T cell nucleus was injected into an enucleated oocyte, and the resulting cell was used to generate embryonic stem (ES) cells. ES cells were injected into recipient blastocysts, and blastocysts implanted into pseudopregnant females. PCR analysis and Sanger sequencing reveal that the rearranged TCR is Vbeta4-D1-J1.6-C1 (trbv2-trbj1.6-trbc1).

H2^d

Allele Symbol: H2^d

Allele Name	d variant
Allele Type	Spontaneous
Allele Synonym(s)
Gene Symbol and Name	H2, histocompatibility-2, MHC
Gene Synonym(s)
Strain of Origin	various
Chromosome	17
General Note	The d variant has been observed in the following strains: DBA/2, DBA/2J BALB/c, BALB/cByJ, BALB/cJ, C57BLKS, NZB.

Ptprc^a

Allele Symbol: Ptprc^a

Allele Name	a variant
Allele Type	Not Applicable
Allele Synonym(s)	CD45.1; Ly5^a; Ptprc^SJL
Gene Symbol and Name	Ptprc, protein tyrosine phosphatase, receptor type, C
Gene Synonym(s)
Site of Expression	Widely expressed on all adaptive and innate immune cells.
Strain of Origin	Not Applicable
Chromosome	1
Molecular Note	Ptprc^a is found in strains SJL/J, STS/A, and DA. Ptprc^b is found in strains C57BL/6, C3H/An, DBA/2, AKR, and many others (J:13367, J:12054, J:12077, J:8603). Twelve nucleotide differences between the a and b alleles have been identified. These base substitutions correspond to five amino-acid changes within the extracellular domain of the encoded protein. These amino-acid differences are clustered in a region that also contains the greatest divergence between mouse and rat sequences (J:22485). Note that the allele designations originally described were reversed in 1987 (J:8603); all publications prior to 1987 show SJL/J, STS/A, and DA as having the b allele and the C57BL/6J group as having the a allele (J:22341).

Tcra^Ln1Bdb

Allele Symbol: Tcra^Ln1Bdb

Allele Name	recombination line 1, Brian Brown
Allele Type	Other (Not Specified)
Allele Synonym(s)
Gene Symbol and Name	Tcra, T cell receptor alpha chain
Gene Synonym(s)
Strain of Origin	(BALB/cJ x C57BL/6J)F1
Chromosome	14
Molecular Note	This allele is generated using somatic cell nuclear transfer (SCNT). First, a lentivirus encoding GFP was injected into (BALB/cJ x C57BL/6J)F1 mice (CB6F1; H-2K). After 2 weeks, GFP-200-208 H-2K tetramer<+> CD8<+> T cells were isolated. The T cell nucleus was injected into an enucleated oocyte, and the resulting cell was used to generate embryonic stem (ES) cells. ES cells were injected into recipient blastocysts, and blastocysts implanted into pseudopregnant females. PCR analysis and Sanger sequencing reveal that the rearranged TCR is Valpha1-J30 (trav7-4, traj30, and trac).

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Research Tools
- Immunology, Inflammation and Autoimmunity Research
  - T Cell Receptor Transgenics
- Genetics Research
  - Tissue/Cell Markers
Immunology, Inflammation and Autoimmunity Research
- Rearranged Antigen-Specific T Cell Receptor Transgenes
  - class I restricted

Genotype: H2^d related

Immunology, Inflammation and Autoimmunity Research
- CD Antigens, Antigen Receptors, and Histocompatibility Markers

Mammalian Phenotype Terms by Genotype

References

Agudo J; Ruzo A; Park ES; Sweeney R; Kana V; Wu M; Zhao Y; Egli D; Merad M; Brown BD. 2015. GFP-specific CD8 T cells enable targeted cell depletion and visualization of T-cell interactions. Nat Biotechnol 33(12):1287-1292PubMed: 26524661 MGI: J:248646

Additional - H2^d related

Additional - Ptprc^a related

Additional - Tcra^Ln1Bdb related

Additional - Tcrb^Ln1Bdb related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Probe:Ptprc End Point-Alternate 1

Breeding Considerations

Jedi (Just EGFP Death Inducing) mice harbor four alleles that are not linked and are expected to segregate independently: the CD45.1 allele (Ptprc^a) on chromosome 1, the two Jedi TCR alleles (Tcrb^Ln1Bdb on chromosome 6 and Tcra^Ln1Bdb on chromosome 14) and the H-2K^d allele (H2^d) on chromosome 17.

When homozygous for CD45.1, heterozygous for the two Jedi TCR alleles and homozygous for H-2K^d, the Jedi mice are viable and fertile. The donating investigator reports that when they breed heterozygous Jedi mice together they obtain the expected number of litters, but their genotyping protocol cannot properly distinguish heterozygous from homozygous to date (August 2018). By November 2019, The Jackson Laboratory has genotyping assays in place for the two Jedi TCR alleles to distinguish heterozygous from homozygous. Additionally, the donating investigator suggests that mice may be immunocompromised when homozygous for the two Jedi TCR alleles.

When maintaining our live colony, mice homozygous for CD45.1, heterozygous or homozygous for the two TCR alleles and homozygous for H-2K^d may be bred together.

Additional Breeding and Husbandry Support

Mating System

Homozygous for Ptprc^a Homozygous for Tcrb^Ln1Bdb Homozygous for Tcra^Ln1Bdb Homozygous for H2^dX Homozygous for Ptprc^a Homozygous for Tcrb^Ln1Bdb Homozygous for Tcra^Ln1Bdb Homozygous for H2^d

Citation
When using the Jedi (Just EGFP Death Inducing) mouse strain in a publication, please cite the originating article(s) and include JAX stock #028062 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX18 (Maximum)

Pricing & Availability

Available

Domestic

International

Live Mouse

Age Genotype Price

weeks

Cryorecovery - Pricing

Service/Product Description Price

Homozygous for Ptprc<a>, homozygous for Tcrb<Ln1Bdb>, homozygous for Tcra<Ln1Bdb>, and homozygous for H2<d>

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Also Known As:Jedi (Just EGFP Death Inducing), Jedi

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Approximate Controls

Additional Information

Selected References

TcrbLn1Bdb

Allele Symbol: TcrbLn1Bdb

H2d

Allele Symbol: H2d

Ptprca

Allele Symbol: Ptprca

TcraLn1Bdb

Allele Symbol: TcraLn1Bdb

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Genotype: H2d related

Mammalian Phenotype Terms by Genotype

References

Additional - H2d related

Additional - Ptprca related

Additional - TcraLn1Bdb related

Additional - TcrbLn1Bdb related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

Live Mouse

Cryorecovery - Pricing

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Tcrb^Ln1Bdb

Allele Symbol: Tcrb^Ln1Bdb

H2^d

Allele Symbol: H2^d

Ptprc^a

Allele Symbol: Ptprc^a

Tcra^Ln1Bdb

Allele Symbol: Tcra^Ln1Bdb

Genotype: H2^d related

Additional - H2^d related

Additional - Ptprc^a related

Additional - Tcra^Ln1Bdb related

Additional - Tcrb^Ln1Bdb related