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
GGATGATGGAGCACCTATGggg
Cas9 target site 2 sequence
Position in the gene exon3
Restriction enzyme (RE) BsiHKAI
PCR primer (forward) tttgtcctgcagGTTCTGGT
PCR primer (reverse) CCCTGAGGACACACACAGAG
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
8b
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)
Genotye Information: