This miR-17~92Δ19 knock-out strain is useful in studies related to the Mir19a and Mir19b-1 seed family, the Mirc1 microRNA cluster, and Myc-driven tumorigenesis and tumor progression.
Andrea Ventura, Memorial Sloan Kettering Cancer Center
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Mirc1 | microRNA cluster 1, including Mir17 through Mir92-1 |
The polycistronic Mirc1, microRNA cluster, or miR-17~92, consists of 6 members: Mir17, microRNA 17; Mir18, microRNA 18; Mir19a, microRNA 19a; Mir20a, microRNA 20a; Mir19b-1, microRNA 19b-1; and Mir92-1, microRNA 92-1. The region of nucleotides 2–7 from the mature miRNA 5´-end is called the miRNA seed sequence, which is essential for target recognition and binding to mRNA. The same seed sequence is often shared amongst miRNAs within the same family.
These miR-17~92Δ19 mice have the miR-19 single seed family (Mir19a and Mir19b-1) knocked out of the Mirc1, microRNA cluster.
No Mir19a or Mir19b-1 miRNAs are detected by RT-qPCR and small RNA sequencing analysis of tail tissue from homozygotes. Mice that are homozygous for the targeted mutation are viable and fertile on the mixed B6;129 background, and slightly smaller than wildtype controls. Mendelian frequency for homozygotes on the C57BL/6J background is lower than expected (5%). 20% of homozygotes on the C57BL/6J background (N7 on the C57BL/6 background) exhibit vertebral fusion. The severity of the phenotype (including perinatal lethality) in homozygotes increases when the mice are backcrossed to C57BL/6.
This strain is part of a miR-17~92 (Mirc1, microRNA cluster 1, including Mir17 through Mir92-1) allelic series:
(Stock No. 027317), (Stock No. 027318),
(Stock No. 027319), (Stock No. 027320), (Stock No. 027321), (Stock No. 027322).
A targeting vector containing a FRT site flanked neo selection cassette was utilized in the construction of this mutant. PCR-mediated mutagenesis was employed to delete
pre-miRNA sequences for Mir19a and Mir19b-1 from the targeting vector.
The construct was electroporated into (C57BL/6 x 129S4/SvJae)F1 derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were tested for germline transmission. Heterozygotes were then bred to beta-Actin-Flpe mice, carrying the Tg(ACTFLPe)9205Dym transgene, on the C57BL/6 background, to excise the FRT flanked NEO cassette. The mice were then backcrossed to C57BL/6J for 10 generations. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
Allele Name | targeted mutation 2.1, Andrea Ventura |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | miR-17~92 delta19 |
Gene Symbol and Name | Mirc1, microRNA cluster 1, including Mir17 through Mir92-1 |
Gene Synonym(s) | |
Strain of Origin | (C57BL/6 x 129S4/SvJae)F1 |
Chromosome | 14 |
Molecular Note | The Mir19a and Mir19b-1 genes were replaced by an FRT-flanked neomycin cassette. Flp-mediated recombination removed the FRT-flanked neo cassette. |
When maintaining a live colony, heterozygous mice may be bred together, to wildtype siblings, or to C57BL/6J inbred mice (Stock No. 000664). The severity of the phenotype (including perinatal lethality) in homozygotes increases when the mice are backcrossed to C57BL/6.
When using the B6.Cg-Mirc1tm2.1Aven/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #027319 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous or wildtype for Mirc1<tm2.1Aven> |
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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. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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