Description

Strain Information

Type Mutant Stock; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Mating System Homozygote x Homozygote         (Female x Male) Species laboratory mouse Generation N?+1F5 (22-OCT-08)   Donating Investigator Brian Harfe,   University of Florida

Description
These mice contain loxP sites on either side of exon 23 of the targeted gene. Mice homozygous for this "Dicer-flox" allele are viable and fertile and exhibit no gross phenotypic or behavioral abnormalities. Expression of the targeted allele is indistinguishable from wildtype despite the presence of an frt-flanked neomycin cassette between exon 23 and the 3' loxP site. Cre-mediated recombination (resulting in deletion of exon 23) in the germline leads to developmental arrest at embryonic day 7.5 (E7.5). Tissue specific deletion results in the loss of microRNA (miRNA) processing. Mutant mice can be used to generate cell/tissue-specific deletions of the endogenous gene for applications in embryonic development, translation, protein processing and miRNA/siRNA regulation of gene expression.

For example, when crossed to a strain expressing Cre recombinase in mesenchyme (see Stock No. 005584), this mutant mouse strain may be useful in studies of limb morphogenesis.

Development
A targeting vector was used to flank exon 23 (encoding most of the second RNaseIII domain) of the endogenous gene with loxP sites. The vector also contained an frt-flanked neomycin resistance cassette between the exon and downstream loxP site. This construct was electroporated into “129” embryonic stem (ES) cells. Correctly targeted cells were injected into C57BL/6J blastocysts and chimeric mice were bred to C57BL/6J for germline transmission. Heterozygous offspring were bred to generate homozygotes. At some point, homozygotes were bred to mutant Gt(ROSA)26Sor mice on an unknown background. Upon arrival at The Jackson Laboratory, mutants will be bred to C57BL/6J mice and then wildtype siblings for multiple generations while selecting for the "Dicer-floxed" allele and against the mutant Gt(ROSA)26Sor allele. After this, mice containing only the "Dicer-floxed" allele will be bred together to create a homozygous colony.

Control Information

	Control
	101043 B6129SF1/J	(approximate)
Considerations for Choosing Controls

Related Strains

Strains carrying   Dicer1^tm1Bdh allele

006366

B6.Cg-Dicer1tm1Bdh/J

View Strains carrying   Dicer1^tm1Bdh     (1 strain)

Additional Web Information

Cre-lox Systems

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

The following phenotype relates to a compound genotype created using this strain.
Contact JAX^® Services jaxservices@jax.org for customized breeding options.

Dicer1^tm1Bdh/Dicer1^tm1Bdh Tg(Prrx1-cre)1Cjt/0

        involves: C57BL/6J * SJL/J   (conditional)

• limbs/digits/tail phenotype
• abnormal forelimb morphology (MGI Ref ID J:100475)
  • at E11 the forelimbs are smaller and at E12.5 the forelimb size resembles that of wild-type forelimbs at E11.5
  • starting at E10.5 increased cell death is seen in the limb mesoderm
• abnormal hindlimb morphology (MGI Ref ID J:100475)
  • the hindlimbs are smaller but not as severely affected as the forelimbs
  • starting at E12.5 increased cell death is seen in the limb mesoderm
• abnormal long bone morphology (MGI Ref ID J:100475)
  • the long bones of the arms and legs appear twisted
• oligodactyly (MGI Ref ID J:100475)
  • forelimbs show loss of digits and some fusion of digits
• skeleton phenotype
• abnormal long bone morphology (MGI Ref ID J:100475)
  • the long bones of the arms and legs appear twisted
• delayed endochondral bone ossification (MGI Ref ID J:100475)
  • bone formation from cartilage is delayed in the long bones of the limbs

View Research Applications

Research Applications

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

Cell Biology Research
Protein Processing
Transcriptional Regulation

Research Tools
Cre-lox System (loxP-flanked Sequences)
Developmental Biology Research (Cre-lox System)
Genetics Research (Mutagenesis and Transgenesis: Cre-lox System)
Genetics Research (Mutagenesis and Transgenesis: transcriptional activation)

Genes & Alleles

Gene & Allele Information

Allele Symbol		Dicer1tm1Bdh
Allele Name		targeted mutation 1, Brian D Harfe
Allele Type		Targeted (Floxed/Frt)
Common Name(s)		Dcrflox; Dicerflox;
Mutation Made By		Brian D. Harfe and Mike McManus,   UFlorida and UCSF
Gene Symbol and Name		Dicer1, Dicer1, Dcr-1 homolog (Drosophila)
Chromosome		12
Gene Common Name(s)		1110006F08Rik; D12Ertd7e; DCR1; DNA segment, Chr 12, ERATO Doi 7, expressed; Dicer; HERNA; KIAA0928; RIKEN cDNA 1110006F08 gene; mKIAA0928;
Molecular Note		A targeting vector was designed to insert loxP sites around the exon that encodes most of the second RNaseIII domain as well as an frt-flanked neo within the loxP-encompassed sequence. Cre-mediated removal of the sequence would result in the loss of 90 amino acids, a loss that does not create a downstream frame-shift or an unstable protein. [MGI Ref ID J:100475]

Genotyping

Genotyping Information

Genotyping Protocols

Dicer1^tm1Bdh, STD PCR, vers. 1

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Harfe BD; McManus MT; Mansfield JH; Hornstein E; Tabin CJ. 2005. The RNaseIII enzyme Dicer is required for morphogenesis but not patterning of the vertebrate limb. Proc Natl Acad Sci U S A 102(31):10898-903. [PubMed: 16040801]  [MGI Ref ID J:100475]

Additional References

Dicer1^tm1Bdh related

Calabrese JM; Seila AC; Yeo GW; Sharp PA. 2007. RNA sequence analysis defines Dicer's role in mouse embryonic stem cells. Proc Natl Acad Sci U S A 104(46):18097-102. [PubMed: 17989215]  [MGI Ref ID J:127467]

Chong MM; Rasmussen JP; Rundensky AY; Littman DR. 2008. The RNAseIII enzyme Drosha is critical in T cells for preventing lethal inflammatory disease. J Exp Med 205(9):2005-17. [PubMed: 18725527]  [MGI Ref ID J:138683]

Cuellar TL; Davis TH; Nelson PT; Loeb GB; Harfe BD; Ullian E; McManus MT. 2008. Dicer loss in striatal neurons produces behavioral and neuroanatomical phenotypes in the absence of neurodegeneration. Proc Natl Acad Sci U S A 105(14):5614-9. [PubMed: 18385371]  [MGI Ref ID J:133780]

Damiani D; Alexander JJ; O'Rourke JR; McManus M; Jadhav AP; Cepko CL; Hauswirth WW; Harfe BD; Strettoi E. 2008. Dicer inactivation leads to progressive functional and structural degeneration of the mouse retina. J Neurosci 28(19):4878-87. [PubMed: 18463241]  [MGI Ref ID J:135193]

Davis TH; Cuellar TL; Koch SM; Barker AJ; Harfe BD; McManus MT; Ullian EM. 2008. Conditional loss of Dicer disrupts cellular and tissue morphogenesis in the cortex and hippocampus. J Neurosci 28(17):4322-30. [PubMed: 18434510]  [MGI Ref ID J:134779]

Harris KS; Zhang Z; McManus MT; Harfe BD; Sun X. 2006. Dicer function is essential for lung epithelium morphogenesis. Proc Natl Acad Sci U S A 103(7):2208-13. [PubMed: 16452165]  [MGI Ref ID J:106072]

Hornstein E; Mansfield JH; Yekta S; Hu JK; Harfe BD; McManus MT; Baskerville S; Bartel DP; Tabin CJ. 2005. The microRNA miR-196 acts upstream of Hoxb8 and Shh in limb development. Nature 438(7068):671-4. [PubMed: 16319892]  [MGI Ref ID J:103424]

Kumar MS; Lu J; Mercer KL; Golub TR; Jacks T. 2007. Impaired microRNA processing enhances cellular transformation and tumorigenesis. Nat Genet 39(5):673-7. [PubMed: 17401365]  [MGI Ref ID J:123829]

Lynn FC; Skewes-Cox P; Kosaka Y; McManus MT; Harfe BD; German MS. 2007. MicroRNA expression is required for pancreatic islet cell genesis in the mouse. Diabetes 56(12):2938-45. [PubMed: 17804764]  [MGI Ref ID J:132315]

Maatouk DM; Loveland KL; McManus MT; Moore K; Harfe BD. 2008. Dicer1 is required for differentiation of the mouse male germline. Biol Reprod 79(4):696-703. [PubMed: 18633141]  [MGI Ref ID J:140872]

O'Rourke JR; Georges SA; Seay HR; Tapscott SJ; McManus MT; Goldhamer DJ; Swanson MS; Harfe BD. 2007. Essential role for Dicer during skeletal muscle development. Dev Biol 311(2):359-68. [PubMed: 17936265]  [MGI Ref ID J:127496]

Zhou X; Jeker LT; Fife BT; Zhu S; Anderson MS; McManus MT; Bluestone JA. 2008. Selective miRNA disruption in T reg cells leads to uncontrolled autoimmunity. J Exp Med 205(9):1983-91. [PubMed: 18725525]  [MGI Ref ID J:138808]

Health & husbandry

Health & Colony Maintenance Information

Animal Health Reports

Room Number           AX11

Colony Maintenance

Breeding & Husbandry	Upon arrival at The Jackson Laboratory, mutants will be bred to B6129F1 (Stock No. 101043) mice for multiple generations while selecting for the "floxed" allele and against the mutant Gt(ROSA)26Sor allele. After this, mice containing only the "floxed" allele will be bred together to create a homozygous colony.
Mating System	Homozygote x Homozygote         (Female x Male)
Diet Information	LabDiet® 5K52/5K67

Purchasing information

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

Pricing

Supply Details

Standard Supply

Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of ~nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within 48 hours of order placement.

Supply Notes

Usually shipped between four and eight weeks of age. This strain is included in the Induced Mutant Resource Colony collection.

Control Information

	Control
	101043 B6129SF1/J	(approximate)
Considerations for Choosing Controls
USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains.
International - Control Pricing Information for Genetically Engineered Mutant Strains.

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.

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Contact Information
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Tel: 800.422.6423 or 207.288.5845
Fax: 207.288.6150
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Terms of Use

Terms of Use

General Terms and Conditions

Effective September 26, 2007: License Requirements for Strains using Cre-lox Technology only apply in Canada, see Licenses for Strains using Cre-lox Technology.

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.”

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