If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO337
Recent progress of this line F1 progeny has been raised
Gene name zgc:85910
Ensembl ID ENSDARG00000056381
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GTCACATCTATTGACTGGGAagg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) Bsr I(ACTGGN|)
PCR primer (forward) AACATCAACGGACTTCTTCAGC
PCR primer (reverse) ATTTCCCGCACCTTCTCAT
Length of expected PCR product (bp) 415
Length of expected RE digestion product (bp) 113+302
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 337
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency Restriction enzyme digestionRE digestion (Bsr I)
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO337.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 33%(1/3)
Wild-type (WT) sequence GTCACATCTATTGACTGGGAAGGACTAATATT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GTCACATCTATTGACTGG-----ACTAATATT(-5bp)
F0-2 GTCACATCTATTGACTGG-AAGGACTAATATT(-1bp) 测序鉴定
Allele name and predicted product 337a
337b
GTCACATCTATTGACTGG-----ACTAATATT; Truncated protein(257aa)
GTCACATCTATTGACTGG-AAGGACTAATATT; Truncated protein(241aa)
突变创建时间 2014-06-03
填表时间 2013/12/18
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information: