RBRC09912

Information1

Image
BRC No.RBRC09912
TypeCRISPR/Cas9 (Transgene)Cartagena
SpeciesMus musculus
Strain nameC57BL/6N-Sik3<em3Iiis>
Former Common nameSik3<Flag,Slp>
H-2 Haplotype
ES Cell line
Background strainC57BL/6NJcl
Appearance
Strain developmentWe induced random point mutations by ethylnitrosourea (ENU) and screened for dominant sleep and wakefulness abnormalities. Through our screening, we established a mutant pedigree, which we termed Sleepy (Slp), with a markedly prolonged sleep time. Whole-exome sequencing identified a heterozygous single nucleotide substitution at the splice donor site for intron 13 of the Sik3 gene. To confirm genetically that the Sik3 splice mutation is the sole cause of the long-sleep phenotype, we introduced the Sik3 exon 13-skipping allele by using conventional knock-in in embryonic stem (ES) cells. Further examine these functions, we introduced a Flag-HA sequence into the Sik3Slp allele using CRISPR/Cas9 technology.
Strain description
Colony maintenance
ReferencesNature, 539, 378-383 (2016). 27806374

Health Report

Examination Date / Room / Rack2018/03/12Room:2-検Rack:D
2018/01/15Room:2-検Rack:D
2017/11/13Room:2-検Rack:D
2017/09/11Room:2-検Rack:D

Gene

Gene info
Gene symbolGene nameChr.Allele symbolAllele namePromoter
Sik3SIK family kinase 39Sik3<em3Iiis>endonuclease-mediated mutation 3, Masashi Yanagisawa and Hiromasa Funato

Gene symbolGene nameChr.Allele symbolAllele namePromoter
FLAGFLAG tag (synthetic)9FLAG

Gene symbolGene nameChr.Allele symbolAllele namePromoter
Frtyeast FRT (flippase recombination target) site9Frt

Gene symbolGene nameChr.Allele symbolAllele namePromoter
HAhuman influenza virus HA (hemagglutinin) tag sequence9HA

Information2

Donor DNApX330-U6-Chimeric_BB-CBh-hSpCas9[human U6 promoter, S. pyogenes gRNA scaffold, human U6 terminator, CMV,chicken hybrid CMV enhancer/chicken beta-actin promotor (CBh), Synthetic DNA 3xFLAG, SV40 nuclear localization signal(NLS), Streptococcus pyogenes SpCas9 (human codon-optimized), bovine GH polyA signal, AAV2 inverted terminal repeat (ITR), f1 phage f1 origin, E. coli Ampicillin resistance gene (AmpR), E. coli pUC origin], mouse Sik3 genomic DNA, Synthetic FLAG tag, Human Influenza HA tag (Hemagglutinin gene), yeast FRT (flipase recombination target) site
Research applicationFLP/frt system
Specific Term and ConditionsIn publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature(s) designated by the DEPOSITOR is requested. Nature, 539, 378-383 (2016).Prior to requesting the BIOLOGICAL RESOURCE, the RECIPIENT must obtain approval from Dr. Masashi Yanagisawa. The availability of the BIOLOGICAL RESOURCE is limited to a RECIPIENT of a not-for profit institution for a not-for-profit research.
DepositorDrs. Hiromasa Funato and Masashi Yanagisawa (University of Tsukuba)
Strain Statusan icon for Frozen embryosFrozen embryos
an icon for Frozen spermFrozen sperm
Strain AvailabilityRecovered litters from cryopreserved embryos (2 to 4 months)
Cryopreserved sperm (within 1 month)
Additional Info.Mouse of the Month Mar 2019
Category I MTA: CRISPR/Cas9 genome edited bioresources ( Japanese / English )

BRC mice in Publications

No Data