If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO961
Recent progress of this line F2 individuals have been indentified
Gene name slkb
Ensembl ID ENSDARG00000012574
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
TGAGCTGGAGGACTACATGGtgg
Cas9 target site 2 sequence
Position in the gene exon2
Restriction enzyme (RE) n/a
PCR primer (forward) GCGTGGTTAAGTATGTTAGCCAC
PCR primer (reverse) GTTGTGATGATCACATGATGCC
Length of expected PCR product (bp) 330
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pSP6-2sNLS-spCas9
Plasmids used for making gRNA pT7-gRNA-ZKO961
Primers used for making gRNA Pr-ZKO 961
Dosage of injected Cas9 mRNA (ng/ul) 50
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 85
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO961.pdf
Mutation efficiency in F0 embryos (%) 30
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 3/10
Wild-type (WT) sequence TGAGCTGGAGGACTACATGGTGG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 TGAGCTGGAGGACT-----GTGG △5
F0-2 TGAGCTGGAGGACTAC-TGGTGG△1
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TGAGCTGGAGGACT-----GTGG △5
F0-2 TGAGCTGGAGGACTAC-TGGTGG△1
Allele name and predicted product 961a
961b
TGAGCTGGAGGACT-----GTGG Truncated protein (84aa)
TGAGCTGGAGGACTAC-TGGTGG Truncated protein (106aa)
突变创建时间 2014-10-15
填表时间 2013/12/19
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information: