If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO338
Recent progress of this line F2 individuals have been indentified
Gene name ankrd37
Ensembl ID ENSDARG00000056376
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GCATCAGTGTGTTGATGGCCagg
Cas9 target site 2 sequence
Position in the gene Exon 3
Restriction enzyme (RE) Sty D4I(|CCNGG)
PCR primer (forward) GCAGCTTCTGCTCTTGCTTATAG
PCR primer (reverse) GCTCATGTATAGGGTGGGTCTTC
Length of expected PCR product (bp) 405
Length of expected RE digestion product (bp) 173+232
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 338
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency Restriction enzyme digestionRE digestion (Sty D4I)
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO338.pdf
Mutation efficiency in F0 embryos (%) 20
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 22%(2/9)
Wild-type (WT) sequence GCATCAGTGTGTTGATGGCCAGG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GCATCAGTGTGTTG--------G(-8bp)
F0-2 GCATCAGTGTG-------CCAGG(-7bp)
Allele name and predicted product 338a
338b
GCATCAGTGTGTTG--------G; Truncated protein(86aa)
GCATCAGTGTG-------CCAGG; Truncated protein(99aa)
突变创建时间 2014-04-10
填表时间 2013/12/18
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information: