If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO10
Recent progress of this line F2 individuals have been indentified
Gene name tmco3
Ensembl ID ENSDARG00000075108
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGCAGAGAAACACCAGCAGG agg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) T7E1
PCR primer (forward) TGGTTGGCCGCTAGTGTATTTT
PCR primer (reverse) TTGTGTTAGTGCACCTGTTTG
Length of expected PCR product (bp) 400
Length of expected RE digestion product (bp) 255+140
Plasmids used for making Cas9 mRNA pSP6-2sNLS-spCas9
Plasmids used for making gRNA pT7-gRNA-ZKOpT7-gRNA-ZKO10
Primers used for making gRNA Pr-ZKO 10
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 91.60
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO10.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 71% (10/14: Number of positive F0 / Total number of F0 screened)
Wild-type (WT) sequence TGGTGGCGGCAGAGAAACACCAGCAGGAGGCG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 TGGTGGCGGCAGAGAAACACCAGCTGAAAGTGAAAAGAGG (+10 bp)
F0-2 TGGTGGCGGCAG--------------GAGGCG (-14 bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TGGTGGCGGCAGAGAAACACCAGCTGAAAGTGAAAAGAGG (+10 bp)
F0-2 TGGTGGCGGCAG--------------GAGGCG (-14 bp)
Allele name and predicted product 10a
10b
GGCAGAGAAACACCAGCTGAAAGTGA Truncated protein (446aa)
TGGTGGCGGCAG--------------GAGGCGTruncated protein (446 aa)
突变创建时间 2014-03-12
填表时间 2013/12/24
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information: