If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO923
Recent progress of this line F1 individuals have been identified
Gene name cryzl1
Ensembl ID ENSDARG00000026902
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGTTGGAAAAACATCACTTT ggg
Cas9 target site 2 sequence
Position in the gene Exon4
Restriction enzyme (RE) T7
PCR primer (forward) TTGTGTTTGTTAGATGTATGAAGAC
PCR primer (reverse) CGTCAATAAGCACGACTGAACAAAG
Length of expected PCR product (bp) 358
Length of expected RE digestion product (bp) 176+182
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 923
Dosage of injected Cas9 mRNA (ng/ul) 500
Dosage of injected gRNA (ng/ul) 200
Survival rate of F0 embryo @ 1dpf (%) 74
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO923.pdf
Mutation efficiency in F0 embryos (%) 16.8
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 20
Wild-type (WT) sequence GTCTGGTTGGAAAAACATCACTTTGGGTCCCA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GTCTGGTTGGAAAAACATCAC+TTTGGGTCCCA (+1bp)
F0-2 GTCTGGTTGGAAAAACATCACT--GGGTCCCA(-2bp)
Allele name and predicted product 923a
923b
GTCTGGTTGGAAAAACATCAC+TTTGGGTCCCA Truncated protein(84aa)
GTCTGGTTGGAAAAACATCACT--GGGTCCCA Truncated protein(82aa)
突变创建时间 2014-09-04
填表时间 2013/12/17
修改时间 2016/8/12
PI/负责人 (工作单位)
Genotype Information: