MicroRNAs (miRNA) are gaining recognition as important post-transcriptional regulators of gene expression. While more than 500 miRNAs have been identified in the mammalian genome, studies addressing their physiological roles are at an early stage and miRNA functions are only now being elucidated. Toward this end, the Sanger Institute is generating a continually expanding collection of MicroRNA Knockout ES cell lines for use by the scientific community through the MMRRC. Additionally, Sanger has created a searchable data portal through which investigators can access additional information such as vector design, PCR results, and gene details. Schematic of MirKO targeting vector and alleles
In the first shipment from Sanger, the MMRRC at UC Davis received 10 96-well plates containing a total of 614 distributable ES cell clones, and will receive additional clones as they are generated. There are approximately 154 different MicroRNA encoding genes represented. The majority of these clones were generated on a JM8A3 background with a small percentage of clones generated on a JM8.F6 background. A subset of these miRNA knockout lines are “cluster” knockouts in which more than one gene has been targeted. The MMRRC catalog pages for these strains indicate the presence of a cluster and strains are named according to the first gene in the cluster.
Requesters should be aware that there are some clones for which it has not been possible to confirm targeting by PCR at both ends. This is most likely due to technical failure of the PCR rather than incorrectly targeted clones; however, investigators are strongly encouraged to confirm targeting by Southern Blot analysis. The MMRRC catalog pages for these strains will note PCR failure where applicable.
The MMRRC holds over 100 lines in this collection. View MMRRC catalog of all Sanger miRNA knockout lines.
Cells from this collection are distributed using the MMRRC Conditions of Use *(COU). Cells are available only to investigators at non-profit institutions.
* The preceding link is to the COU text only; the COU user form will be provided after order submission.
For additional assistance with these clones, please contact the MMRRC at UC Davis at (530) 754-3290 or email firstname.lastname@example.org.
Information here is excerpted from the following publication: