If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO334
Recent progress of this line F2 individuals have been indentified
Gene name si:ch211-197n1.2
Ensembl ID ENSDARG00000042937
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGAGCTCCCAGTCTCTTCCagg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) Sty D4I(|CCNGG)
PCR primer (forward) TGATGGTGAAGACCGACATTG
PCR primer (reverse) CAAGTTCCAGAACAGGAGAAATTC
Length of expected PCR product (bp) 468
Length of expected RE digestion product (bp) 285+75+108
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 334
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 70
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/ZKO334.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 60%(3/5)
Wild-type (WT) sequence GGAGCTCCCAGTCTCTTCC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGAGCTCCCAGTC---TCC(-3bp)
F0-2 GGAGCTC----------CC(-10bp)测序鉴定
Allele name and predicted product 334a
334b
GGAGCTCCCAGTC---TCC; Truncated protein(889bp)
GGAGCTC----------CC; Truncated protein(547aa)
突变创建时间 2014-04-17
填表时间 2013/12/18
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information: