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微信小章| 产品名称： | Dicer1-/- |
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| 商品货号： | TS189613 |
| Organism： | Mus musculus, mouse |
| Tissue： | Derived from: mesenchyme |
| Cell Type： | mesenchymal stem cell |
| Product Format： | frozen 1.0 mL |
| Morphology： | mesenchymal-like |
| Culture Properties： | adherent |
| Biosafety Level： | 2 xa0Cells contain SV40 viral DNA sequences
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Age： | 13 month |
| Gender： | female |
| Applications： | Dicer1-/- mesenchymal stem cells (ATCC® CRL-3221™) lack expression of the Dicer1 protein. Dicer1-/- cells can be used in conjunction with the matching Dicer1f/f cells (ATCC® CRL-3220™) to study the role of Dicer and miRNA in cell biology. |
| Storage Conditions： | liquid nitrogen vapor phase |
| Images： |  |
| Derivation： | The cells were first derived as primary cultures from a Dicer1f/f mouse with loxP sites present on either side of exon 23 of the Dicer1 gene. The primary culture was then infected with ecotropic virus encoding SV40 large T-Antigen, and immortalized. The immortalized MSCs then were infected with Adeno-Cre-GFP to introduce Dicer gene knockout by Cre/loxP- mediated recombination. The cells were sorted for GFP. Clones were then isolated and genotyped. |
| Genes Expressed： | functional Dicer1 not expressed |
| Comments： | MicroRNAs (miRNAs) are a new class of small noncoding RNAs that post-transcriptionally regulate the expression of target mRNA transcripts. Dicer is an RNase III family endoribonuclease that has the biofunction of processing miRNAs. Loss of Dicer1 leads to a dramatic decrease in levels of cellular microRNA, allowing for investigations into the roles of microRNAs in various cellular functions. CRL-3221 the Dicer1-/- cell line is a model of homozygous Dicer1-deletion in murine mesenchymal stem cells established from an adult Dicer1f/f mouse and immortalized in vitro. CRL-3220 is the Dicer1f/f immortalized mouse mesenchymal stem cell line that can be paired with CRL-3221 and used as a control. |
| Complete Growth Medium： | Alpha minimum essential medium with ribonucleosides and deoxyribonucleosides, 90%; fetal bovine serum, 10%
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| Subculturing： | Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Medium Renewal: 2 to 3 times per week Seeding Density: 1.5 x 104 to 3.0 x 104 cells/cm2 |
| Cryopreservation： | Freeze Medium: Fetal Bovine Serum (FBS), 92%; DMSO, 8% Storage Temperature: liquid nitrogen vapor phase |
| Culture Conditions： | Temperature:37°C Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Volume： | 1.0 mL |
| Name of Depositor： | A Gurtan (P Sharps lab) |
| Year of Origin： | 2009 |
| References： | Ravi A, et al. Proliferation and tumorigenesis of a murine sarcoma cell line in the absence of DICER1. Cancer Cell 21(6): 848-855, 2012. PubMed: 22698408 Gurtan AM, et al. In vivo structure-function analysis of human Dicer reveals directional processing of precursor miRNAs. RNA 18: 1116-1122, 2012. PubMed: 22546613 |