If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1090
Recent progress of this line F2 generation has been raised
Gene name tektin 4
Ensembl ID ENSDARG00000028899
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGCGCTCCGGTAACCCGCGGtgg
Cas9 target site 2 sequence
Position in the gene exon1
Restriction enzyme (RE) n/a
PCR primer (forward) CAAACTAACCGTTCGTCATC
PCR primer (reverse) TGCGCTGAGCCGCTCAAT
Length of expected PCR product (bp) 475
Length of expected RE digestion product (bp) n/a
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1090
Dosage of injected Cas9 mRNA (ng/ul) 200
Dosage of injected gRNA (ng/ul) 200
Survival rate of F0 embryo @ 1dpf (%) 50
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1090.pdf
Mutation efficiency in F0 embryos (%) 25
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 40% (2/5)
Wild-type (WT) sequence CGGTTTGACCACCGCGGGTTACCGGAGCGCCAAATACACCC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 CCGGTTTGACCACCG----------GAGCGCCAAATACACCCCC
F0-2 CCGGTTTGACCACC-------ACCG----GCCAAATACACCCCC
Allele name and predicted product 1090a
1090b
CCGGTTTGACCACCG----------GAGCGCCAAATACACCCCC Truncated protein (65aa)
CCGGTTTGACCACC-------ACCG----GCCAAATACACCCCC Truncated protein (60aa)
突变创建时间 2014-03-01
填表时间 2013/12/20
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information: