If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1024
Recent progress of this line F2 generation has been raised
Gene name cnn1a
Ensembl ID ENSDARG00000017193
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGGTGCACTGAACCTGAGTGtgg
Cas9 target site 2 sequence
Position in the gene exon4
Restriction enzyme (RE) AvaII
PCR primer (forward) ACGCATCTAACGAATGGGCT
PCR primer (reverse) GCTACATGTTGCAGTGTTTTCT
Length of expected PCR product (bp) 310
Length of expected RE digestion product (bp) 75bp,172bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 25
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1024.pdf
Mutation efficiency in F0 embryos (%) 35
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0 20
Wild-type (WT) sequence TATTTTTGAGGCCAATGATCTTTTCGAGGACATGAACCACACTCAGGTTCAGTGCACCCTCGTCGCTTTGGCTGGACTG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 TATTTTTGAGGCCAATG..CTTTTCGAGGACATGAACCACACTCAGGTTCAGTGCACCCTCGTCGCTTTGGCTGGACTG ZKO1024a: -2 bp deletion
F0-2 TATTTTTGAGGCCAATGATCTTTTCGAGGACATGAACCAGGTTCAGTGCACCCTCGTCGCTTTGGCTGGACTG ZKO1024b: -6bp
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TATTTTTGAGGCCAATG..CTTTTCGAGGACATGAACCACACTCAGGTTCAGTGCACCCTCGTCGCTTTGGCTGGACTG ZKO1024a: -2 bp deletion
F0-2 TATTTTTGAGGCCAATGATCTTTTCGAGGACATGAACCAGGTTCAGTGCACCCTCGTCGCTTTGGCTGGACTG ZKO1024b: -6 bp deletion
Allele name and predicted product 1024a
1024b
TATTTTTGAGGCCAATG..CTTTTCGAGGACATGAACCACACTCAGGTTCAGTGCACCCTCGTCGCTTTGGCTGGACTG(131)turncate protein
TATTTTTGAGGCCAATGATCTTTTCGAGGACATGAACCAGGTTCAGTGCACCCTCGTCGCTTTGGCTGGACTG(-2aa)
突变创建时间 2013-08-12
填表时间 2013/12/23
修改时间 2016/10/24
PI/负责人 (工作单位)
Genotype Information: