Description

Strain Information

Type Mutant Stock; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Species laboratory mouse Generation F?pN1   Donating Investigator Robert Williams,   Duke University School of Medicine

Description
Mice homozygous for the Cycs^tm1Wlm targeted-mutant allele die in utero by embryonic day 10.5, but cell lines established from early Cycs-null embryos are viable under conditions that compensate for defective oxidative phosphorylation. Cells lacking cytochrome c show reduced caspase 3 activation, and are resistant to the proapoptotic effects of UV irradiation, serum withdrawal, and staurosporine. Cells lacking cytochrome c, however, do demonstrate an increased sensitivity to cell death signals triggered by tumor necrosis factor, alpha. Heterozygous mice are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.

Development
A targeting vector containing neomycin resistance and thymidine kinase genes was used to disrupt the entire gene and flanking sequences. The construct was electroporated into 129-derived KG-1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric animals were crossed to C57BL/6 mice.

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

Cycs^tm1Wlm/Cycs^tm1Wlm

        involves: 129X1/SvJ * C57BL/6

• lethality-prenatal/perinatal
• embryonic lethality during organogenesis (MGI Ref ID J:62276)
  • homozygous null embryos died by midgestation; no viable embryos were found after E10.5
• cellular phenotype
• abnormal aerobic energy metabolism (MGI Ref ID J:62276)
  • cells derived from E8.5 mutant embryos showed respiratory insufficiency and enhanced anaerobic glycolysis
  • cell viability and proliferation improved by supplementing the medium with uridine and pyruvate
• abnormal apoptosis (MGI Ref ID J:62276)
  • mutant cells showed reduced caspase-3 activation; caspase-3 activating activity could be rescued by exogenous cytochrome c
  • mutants cells were resistant to the proapoptotic effects of UV irradiation, serum withdrawal, or staurosporine
  • mutants cells showed increased sensitivity to cell death signals triggered by TNF, both in the presence or absence of cycloheximide
• embryogenesis phenotype
• abnormal extraembryonic tissue morphology (MGI Ref ID J:62276)
  • at E9.5, mutant embryos were enclosed in a ball-like structure formed by the yolk sac and the amnion
• embryonic growth retardation (MGI Ref ID J:62276)
  • up to E9.5, development appeared to progress in a relatively normal but delayed manner
  • at E9.5, mutant embryos displayed a primitive heart tube, somites and an allantois, features consistent with ~E8 in wild-type
• reduced embryo size (MGI Ref ID J:62276)
  • by E8.5, homozygous null embryos had a strikingly reduced size
• growth/size phenotype
• embryonic growth retardation (MGI Ref ID J:62276)
  • up to E9.5, development appeared to progress in a relatively normal but delayed manner
  • at E9.5, mutant embryos displayed a primitive heart tube, somites and an allantois, features consistent with ~E8 in wild-type
• reduced embryo size (MGI Ref ID J:62276)
  • by E8.5, homozygous null embryos had a strikingly reduced size

View Research Applications

Research Applications

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

Cycs^tm1Wlm related

Apoptosis Research
Death Receptors
Endogenous Regulators
Extracellular Modulators

Cancer Research
Genes Regulating Growth and Proliferation

Cardiovascular Research
Heart Abnormalities

Cell Biology Research
DNA Damage Response
Genes Regulating Growth and Proliferation

Developmental Biology Research
Embryonic Lethality (Homozygous)
Growth Defects

Research Tools
Apoptosis Research

Genes & Alleles

Gene & Allele Information

Allele Symbol		Cycstm1Wlm
Allele Name		targeted mutation 1, Robert Sanders Williams
Allele Type		Targeted (knock-out)
Common Name(s)		Cyt c-;
Mutation Made By		Robert Williams,   Duke University School of Medicine
Strain of Origin		129S6/SvEvTac
ES Cell Line Name		KG1/KG-1
ES Cell Line Strain		129S6/SvEvTac
Gene Symbol and Name		Cycs, cytochrome c, somatic
Chromosome		6
Gene Common Name(s)		CYC; CYCSA; HCS; MGC93634;
Molecular Note		A neomycin resistance cassette replaced the two protein coding exons. Immunoblot analysis did not detect endogenous protein in cells cultured from E8-E9 embryos of homozygous mutant mice. [MGI Ref ID J:62276]

Genotyping

Genotyping Information

Genotyping Protocols

Cycs^tm1Wlm, STD PCR, vers. 1

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Li K; Li Y; Shelton JM; Richardson JA; Spencer E; Chen ZJ; Wang X; Williams RS. 2000. Cytochrome c deficiency causes embryonic lethality and attenuates stress-induced apoptosis. Cell 101(4):389-99. [PubMed: 10830166]  [MGI Ref ID J:62276]

Additional References

Cycs^tm1Wlm related

Mansfield KD; Guzy RD; Pan Y; Young RM; Cash TP; Schumacker PT; Simon MC. 2005. Mitochondrial dysfunction resulting from loss of cytochrome c impairs cellular oxygen sensing and hypoxic HIF-alpha activation. Cell Metab 1(6):393-9. [PubMed: 16054088]  [MGI Ref ID J:129813]

Vempati UD; Diaz F; Barrientos A; Narisawa S; Mian AM; Millan JL; Boise LH; Moraes CT. 2007. Role of cytochrome C in apoptosis: increased sensitivity to tumor necrosis factor alpha is associated with respiratory defects but not with lack of cytochrome C release. Mol Cell Biol 27(5):1771-83. [PubMed: 17210651]  [MGI Ref ID J:118859]

Health & husbandry

Health & Colony Maintenance Information

Colony Maintenance

Breeding & Husbandry	This strain originated on a B6;129 background. The donating investigator maintains this strain by mating heterozygotes.

Purchasing information

Pricing, Supply Level & Notes, Controls, General Terms & Conditions

Pricing

Supply Details

Standard Supply

Repository-Cryopreserved. Must Be Recovered. Please refer to pricing and supply notes for further information.

Supply Notes

Cryorecovery - Standard. The recovery process begins when a signed agreement form is returned to the Customer Service Department after order placement. Although results vary by strain, at least two males and two females (two pairs) will be provided, typically within 15 weeks of our receipt of the signed agreement form. If the first recovery attempt is unsuccessful or only one pair is recovered, a second recovery will be done, extending the delivery time to approximately 25 weeks. At least one member of each pair will be of known genotype and will carry the mutation if it is a mutant strain. Please note that pairs may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation of the strain. Mating schemes are sometimes modified for successful cryopreservation. Price represents a repository maintenance fee, which includes the cost of recovery of the strain from the cryopreservation resource and the periodic replacement of the frozen embryos used for recovery. Cryorecovery to establish a Dedicated Supply for greater quantities of mice. One to two pairs will be recovered to establish a Dedicated Supply of mice. Price by quotation. For more information on Dedicated Supply, please contact JAX® Services, Tel: 1-800-422-6423 or 1-207-288-5845. This strain is included in the Induced Mutant Resource Colony collection.

General Terms and Conditions

See Terms of Use

The Jackson Laboratory's Genotype Promise

The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX^® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX^® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX^® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.

---

Ordering and Purchasing Information

      Purchasing Information
      JAX^® Mice Orders
      Surgical Services

Contact Information
Orders & Technical Support
Tel: 800.422.6423 or 207.288.5845
Fax: 207.288.6150
Technical Support Email Form

Terms of Use

Terms of Use

General Terms and Conditions

For Licensing and Use Restrictions view the link(s) below:
- Use of MICE by companies or for-profit entities requires a license prior to shipping.

Contact information

General inquiries

Contracts Administration

phone:	207-288-6470
fax:	207-288-6655

JAX^® Mice & Services Conditions of Use

“Each recipient institution, including its employees and other researchers under its control (RECIPIENT), of mice or services using mice from The Jackson Laboratory (TJL) agrees that such mice, descendants of those mice derived by inbreeding or crossbreeding, including unmodified derivatives of those mice or their descendants (“MICE”) shall not be: (i) used for any purpose other than the internal research of the RECIPIENT, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services with respect to MICE. Acceptance of MICE from TJL shall be deemed agreement by RECIPIENT to these conditions, and departure from these conditions requires The Jackson Laboratory’s prior written authorization.”

In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of MICE, products or services, The Jackson Laboratory will, at its option, provide credit or replacement for the MICE or product received or the services provided.

No Liability

In no event shall The Jackson Laboratory, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, products or services, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of The Jackson Laboratory, its agents or employees. In purchasing or receiving MICE, products or services from The Jackson Laboratory, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges The Jackson Laboratory from all such causes of action or damages, and further agrees to defend and indemnify The Jackson Laboratory from any costs or damages arising out of any third party claims.

MICE and biological materials are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.

The foregoing represents the General Terms and Conditions applicable to The Jackson Laboratory’s MICE, products and services. In addition, special terms and conditions of sale of certain MICE, products and services may be set forth separately in The Jackson Laboratory web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, products and services by The Jackson Laboratory, and by its licensees and distributors.

Acceptance of delivery of MICE, products or services shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on The Jackson Laboratory, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, products services by The Jackson Laboratory.