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Strain Detail Sheet

Strain Name:
Keck MirKO ES cell line Mir182/ Mir-96/ Mir-183
Stock Number:
036877-UCD
Citation ID:
RRID:MMRRC_036877-UCD
Other Names:
miR-182/ mir-96/ mir-183
Major Collection:
Keck MirKO

Gene Information

Gene Details

Genetic Alterations
A targeting vector was designed to insert an FRT site followed by a lacZ gene, a loxP site, a neomycin cassette, an FRT site and a loxP site (FRT-lacZ-loxP-Neo-FRT-loxP) upstream of the miR stem loop, with one loxP site placed immediately downstream of the miR stem loop.
Genotype Determination
ES Cell Line
E14TG2a.4 derived from 129P2/OlaHsd

Strain Description

Strain of Origin
129P2/OlaHsd

Related Protocols

Strain Origin

Donor
Michael McManus, Ph.D., University of California, San Francisco, W.M.Keck Center for Noncoding RNAs
Primary Reference
Park CY, Jeker LT, Carver-Moore K, Oh A, Liu HJ, Cameron R, Richards H, Li Z, Adler D, Yoshinaga Y, Martinez M, Nefadov M, Abbas AK, Weiss A, Lanier LL, de Jong PJ, Bluestone JA, Srivastava D, McManus MT. A Resource for the Conditional Ablation of microRNAs in the Mouse. Cell Rep. 2012 Apr 19;1(4):385-91. (Medline PMID: 22570807).

Colony and Husbandry Information

Special Considerations

ES Cell Sex and Genotype: These ES Cells are expected to be male (XY) and heterozygous/hemizygous for the mutation.

Health Status Report

Requested cell lines are tested by PCR analysis for mycoplasma spp. and Murine Parvoviruses (MPV) upon completion of expansion and prior to shipment.

Mice derived from requested cell lines via Microinjection services will be pathogen tested prior to delivery of mice and health reports will be provided. If you require additional health status information prior to requesting microinjection please email mmrrc@ucdavis.edu.

Order Request Information

Availability Level

MMRRC Testing of the Sanger and Keck MirKO cell lines includes genetic verification at the 3' and/or 5' ends, vector copy number = 1, presence or loss of Y chromosome, and mycoplasma and MPV testing. Based on the average germline rates, we recommend requesting and injecting 2-3 clones per targeted allele. All mutant ES cell lines are heterozygous for the mutation, meaning they carry one modified allele and one wild type allele. MMRRC also recommends performing chromosome counts or karyotyping prior to injection of clones.

Estimated timelines: The repository was sent only one or two vials per clone/cell line from the production center. Processing ES cells (including genetic verification) prior to shipping or injection requires 6-8 weeks. Chromosome counting (to determine % euploid cells) requires an additional 4-8 weeks. Microinjection requires an additional 11-15 weeks. Germline transmission testing (breeding of chimera) requires an additional 8-16 weeks. Germline transmission is not guaranteed.

Conditions of Distribution

Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.

The donor or their institution limits the distribution to non-profit institutions only.

Fees

Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.

Click button to Request this one strain. (Use the MMRRC Catalog Search to request more than one strain.)
MMRRC Item # Description Distribution Fee / Unit (US $)
*Shipping & Handling not included*		 Units Notes
036877-UCD-CELL Keck MirKO ES cell line Mir182/ Mir-96/ Mir-183 $1,517.22 / Non-Profit Vial Includes genetic confirmation
additional products Additional Clones $681.34 / Non-Profit per clone
available service Chromosome counts $385.17 / Non-Profit per clone To determine the percentage of euploid metaphase chromosomes in the targeted ES cells. At least 20 spreads are counted.
additional products Extra vial of cells for the same clone $462.69 / Non-Profit per clone
available service Basic microinjection service $11,114.27 / Non-Profit per cell line In addition to the per mutant ES cell line fee (see above); chimeras not guaranteed. Injection of up to 3 clones.
additional products Feeder cells - Irradiated $148.67 / Non-Profit per cell line
015890-UCD-CTRL BayGenomics High Passage Parental cell line ES Parental cell line E14Tg2a.4 (suitable for DNA extraction, control purposes; not for germline transmission) $148.67 / Non-Profit Vial Suitable for DNA extraction, control purposes but not for germline transmission.
015890-UCD-CELL BayGenomics Low Passage Parental cell line ES Parental cell line E14Tg2a.4 (suitable for electroporation and germline transmission) $740.70 / Non-Profit Vial Suitable for a single electroporation and germline transmission.
015890-UCD-ULTRA BayGenomics Ultra-low Passage Parental cell line ES Parental cell line E14Tg2a.4 (suitable for multiple electroporations and germline transmission) $2,059.70 / Non-Profit Vial Suitable for a multiple electroporation and germline transmission.
To request material from the MMRRC: Please fill out our on-line request form (accessible from the catalog search results page, or click the Request this Strain button in the fees section). If you have questions or need assistance completing this form, you may call Customer Service at (800) 910-2291 (in USA or Canada) or (530) 757-5710 (international calls). Before you call, please have with you: the MMRRC item number, quantity needed, Bill-to and Ship-to contact information.