If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO8
Recent progress of this line F2 individuals have been indentified
Gene name adprhl1
Ensembl ID ENSDARG00000041589
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
Cas9 target site 2 sequence
Position in the gene exon3
Restriction enzyme (RE) BsiHKAI
PCR primer (forward) tttgtcctgcagGTTCTGGT
Length of expected PCR product (bp) 227
Length of expected RE digestion product (bp) 130+90
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO
Dosage of injected Cas9 mRNA (ng/ul) 500
Dosage of injected gRNA (ng/ul) 100
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency Restriction enzyme digestionBsiHKAI digestion
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO8.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 20%
Wild-type (WT) sequence CCACAACCACCCCATAGgtgctccatcatcctccccctcc
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 CCACAACCAC----------ctccatcatcctccccctcc (-10bp) CCACAACCACCCCATcAtCctcctccatcatcctccccctcc (+2bp)
F0-2 C-------A--Tc--A-----tccatcatcctccccctcc (-16bp) CCACAACCACCCCAT-----------catcctccccctcc (-11bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 18%(9/48) C-------A--Tc--A-----tccatcatcctccccctcc -16bp
F0-2 8%(4/48) CCACAACCAC----------ctccatcatcctccccctcc (-10bp)
Allele name and predicted product 8a
C-------A--Tc--A-----tccatcatcctccccctcc change splicing
突变创建时间 2014-01-02
填表时间 2013/12/17
修改时间 2016/12/30
PI/负责人 (工作单位)
Genotype Information: