title: yKI1
organism: Mus musculus
description: Protocols: Knock-in in the Cebpb locus that replaces the wt Cebpb-mRNA sequence for a mutated version that contains a mutation in the AUG-initiation codon of the cis-regulatory upstream open reading frame (uORF), abrogating uORF function. The exact mutation is wt AGCATGCCT -> delta-uORF AGAATTCCT (introducing EcoRI site). This mutation results in deficiency of uORF-dependent translation into the C/EBPbeta-LIP protein isoform, leaving translation into the C/EBPbeta-LAP protein isoforms unaffected. Liver tissue from young (5 months) and old (20 months) wt and C/EBPβΔuORF mice (from six individuals per group) was homogenized on ice with a motor driven pellet pestle (Kontes) in the presence of QIAzol reagent (Qiagen) and total RNA was isolated as described in Zidek et al., 2015. RNA was isolated from tissue samples using the RNeasy® Lipid Tissue Mini Kit (QIAGEN). After incubation for 30 min at 37°C with RQ1 RNase‐Free DNase (Promega), the RNA was further purified with the RNeasy® Plus Mini Kit (QIAGEN) according to the manufacture's protocol starting at step 4. Preparation of the sequencing libraries was performed using the TruSeq Sample Preparation V2 Kit (Illumina) according to the manufacturer's instructions.
attributes.tag: ENA first public
attributes.value: 2019-01-16
attributes.tag: ENA last update
attributes.value: 2018-01-31
attributes.tag: External Id
attributes.value: SAMEA104556852
attributes.tag: INSDC center alias
attributes.value: European Research Institute for the Biology of Ageing, University of Groningen, UMC Groningen, The Netherlands
attributes.tag: INSDC center name
attributes.value: European Research Institute for the Biology of Ageing, University of Groningen, UMC Groningen, The Netherlands
attributes.tag: INSDC first public
attributes.value: 2019-01-16T17:02:03Z
attributes.tag: INSDC last update
attributes.value: 2018-01-31T10:18:11Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6435:yKI1
attributes.tag: age
attributes.value: 4
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: genotype
attributes.value: C/EBP uORF expression
attributes.tag: organism part
attributes.value: liver
attributes.tag: sample name
attributes.value: E-MTAB-6435:yKI1
attributes.tag: sex
attributes.value: female
attributes.tag: strain
attributes.value: C57BL/6