Overview

Also Known As:12EN ESYT1/ESYT2 conditional KO

Esyt1/Esyt2 conditional mice possess recombination cassettes that allow for the development of different conditional, KO, and KI lines. These mice are useful when studying Ca²⁺ signaling at endoplasmic reticulum-plasma membrane junctions.

Donating Investigator

Dr. Thomas C. Sudhof, Stanford University School of Medicine

Genetic overview

Genetic Background	Generation

Esyt2^tm1Sud

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), No functional change)	Esyt2	extended synaptotagmin-like protein 2

Esyt1^tm1Sud

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), No functional change)	Esyt1	extended synaptotagmin-like protein 1

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Research Applications

Research Tools
Cell Biology Research

View All Research Applications

Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

Esyt1/Esyt2 conditional mice possess recombination cassettes that allow for the development of different conditional and KO lines. Esyt1 and Esyt2 encode membrane proteins that are involved with the formation of junctions between endoplasmic reticulum (ER) and plasma membrane (PM), as well as the Ca²⁺ signaling and lipid trafficking at ER-PM junctions. Each allele contains a mutation in exon 10 that changes four aspartate residues into alanine residues (D4A). The aspartate residues are required for calcium binding of the ESYT1 and ESYT2 proteins, therefore when mutated into alanines the proteins should not be able to bind calcium. Mice that are homozygous for both alleles are viable and fertile.

When bred to mice expressing FLP Recombinase, recombination of frt sites should result in a strain with loxP sites flanking exon 10 of the Esyt1 and Esyt2 genes. Subsequent breeding to a strain expressing Cre Recombinase can be expected to result in offspring with the floxed exons deleted, creating a double KO strain.

When bred to a mouse expressing FLP Recombinase, recombination of F3 sites is designed to remove the endogenous exons 10 of both Esyt1 and Esyt2 genes, and only express the mutated D4A version of each exon.

Development

A targeting vector was designed to insert an F3 site followed by an attB site and a loxP site upstream of exon 10 of the extended synaptotagmin-like protein 1 (Esyt1) gene. Inserted downstream of exon 10, from 5’ to 3’, are a second loxP site, an attP site, a frt site, a neomycin resistance cassette, a second F3 site, and a mutated exon 10 with four aspartate residues mutated into alanine residues (D4A) followed by a second frt site. The construct was electroporated into C57BL/6-derived B6-3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred to C57BL/6 mice.

Another targeting vector was designed to insert an F3 site followed by a loxP site upstream of exon 10 of the extended synaptotagmin-like protein 2 (Esyt2) gene. Inserted downstream of exon 10, from 5’ to 3’, are a second loxP site, a frt site, a neomycin resistance cassette, a second F3 site, a loxP2272 site, and a mutated exon 10 with four aspartate residues mutated into alanine residues (D4A) followed by a second loxP2272 site and a second frt site. The construct was electroporated into C57BL/6-derived B6-3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred to C57BL/6 mice.

Mice from each line were bred together to generate these double conditional mice. Mice were maintained on a mixed B6;129sv background. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.

Control Suggestions

Approximate Controls

101045 B6129SF2/J

Additional Information

Considerations for Choosing Controls

Genetics

Esyt2^tm1Sud

Allele Symbol: Esyt2^tm1Sud

Allele Name	targeted mutation 1, Thomas C Sudhof
Allele Type	Targeted (Conditional ready (e.g. floxed), No functional change)
Allele Synonym(s)	Esyt2^tm1Sud; targeted mutation 1, Thomas C Sudhof
Gene Symbol and Name	Esyt2, extended synaptotagmin-like protein 2
Gene Synonym(s)	CHR2SYT; D12Ertd551e; DNA segment, Chr 12, ERATO Doi 551, expressed; FAM62B; 2410017M09Rik; 4921504I16Rik; E-Syt2; 4921504I16Rik; Fam62b; family with sequence similarity 62 (C2 domain containing), member B; RGD1565705; RIKEN cDNA 2410017M09 gene; D12Ertd551e; RIKEN cDNA 4921504I16 gene; 2410017M09Rik; Fam62b
Strain of Origin	C57BL/6NTac
Chromosome	12
Molecular Note	A targeting vector is designed to insert an F3 site followed by a loxP site upstream of exon 10. Inserted downstream of exon 10, from 5' to 3' , are a second loxP site, a FRT site, a neomycin resistance cassette, a second F3 site, a loxP2272 site, and a mutated exon 10 with four aspartate residues mutated into alanine residues (D4A) followed by a second loxP2272 site and a second FRT site.

Esyt1^tm1Sud

Allele Symbol: Esyt1^tm1Sud

Allele Name	targeted mutation 1, Thomas C Sudhof
Allele Type	Targeted (Conditional ready (e.g. floxed), No functional change)
Allele Synonym(s)	Esyt1^tm1Sud; targeted mutation 1, Thomas C Sudhof
Gene Symbol and Name	Esyt1, extended synaptotagmin-like protein 1
Gene Synonym(s)	Fam62a; FAM62A; family with sequence similarity 62 (C2 domain containing), member A; vp115; Mbc2; membrane bound C2 domain containing protein; MBC2; Mbc2; Fam62a
Strain of Origin	C57BL/6NTac
Chromosome	10
Molecular Note	A targeting vector was designed to insert an F3 site followed by an attB site and a loxP site upstream of exon 10. Inserted downstream of exon 10, from 5'to 3' are a second loxP site, an attP site, a FRT site, a neomycin resistance cassette, a second F3 site, and a mutated exon 10 with four aspartate residues mutated into alanine residues (D4A) followed by a second FRT site.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

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

Research Tools
- Cre-lox System
  - loxP-flanked Sequences
- FLP-FRT System
  - FRT-flanked Sequences
Cell Biology Research
- Signal Transduction
- Channel and Transporter Defects
  - calcium

Mammalian Phenotype Terms by Genotype

The following phenotype relates to a compound genotype created using this strain

Genotype: Esyt1^tm1Sud/Esyt1^tm1Sud Esyt2^tm1Sud/Esyt2^tm1Sud
B6;129-Esyt1 Esyt2/J

normal phenotype

no abnormal phenotype detected
- mice are viable and fertile

References

Additional - Esyt1^tm1Sud related

Additional - Esyt2^tm1Sud related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Esyt1alternate2
- Standard PCR:Esyt2
- Sanger sequencing:Esyt2(D4A)
- Sanger sequencing:Esyt1(D4A)
- Standard PCR:Generic Neo
- Probe:Generic Neo
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, double homozygous mice may be bred together.
- Additional Breeding and Husbandry Support
Citation
When using the 12EN ESYT1/ESYT2 conditional KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #027583 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

Cryo Recovery

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Heterozygous or wildtype for Esyt1<tm1Sud>, Heterozygous or wildtype for Esyt2<tm1Sud>

Related Products and Services

Frozen Mouse Embryo	B6;129-Esyt1<tm1Sud> Esyt2<tm1Sud>/J Frozen Embryos	$2595.00
Frozen Mouse Embryo	B6;129-Esyt1<tm1Sud> Esyt2<tm1Sud>/J Frozen Embryos	$2595.00
Frozen Mouse Embryo	B6;129-Esyt1<tm1Sud> Esyt2<tm1Sud>/J Frozen Embryos	$3373.50
Frozen Mouse Embryo	B6;129-Esyt1<tm1Sud> Esyt2<tm1Sud>/J Frozen Embryos	$3373.50

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Also Known As:12EN ESYT1/ESYT2 conditional KO

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Approximate Controls

Additional Information

Esyt2tm1Sud

Allele Symbol: Esyt2tm1Sud

Esyt1tm1Sud

Allele Symbol: Esyt1tm1Sud

Disease Terms

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Esyt1tm1Sud/Esyt1tm1Sud Esyt2tm1Sud/Esyt2tm1SudB6;129-Esyt1 Esyt2/J

normal phenotype

References

Additional - Esyt1tm1Sud related

Additional - Esyt2tm1Sud related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Related Products and Services

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

Esyt2^tm1Sud

Allele Symbol: Esyt2^tm1Sud

Esyt1^tm1Sud

Allele Symbol: Esyt1^tm1Sud

Genotype: Esyt1^tm1Sud/Esyt1^tm1Sud Esyt2^tm1Sud/Esyt2^tm1Sud
B6;129-Esyt1 Esyt2/J

Additional - Esyt1^tm1Sud related

Additional - Esyt2^tm1Sud related