If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1021
Recent progress of this line F2 individuals have been indentified
Gene name best2
Ensembl ID ENSDARG00000079163
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGCCTTCTTCGCCATGTACAcgg
Cas9 target site 2 sequence
Position in the gene Exon 1
Restriction enzyme (RE) NlaIII(CATG)
PCR primer (forward) CTTCTGCGGCTTCTCCAAACTCCTG
PCR primer (reverse) CATGCACACACCTGTACGTG
Length of expected PCR product (bp) 125
Length of expected RE digestion product (bp) 89,37
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1021
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 87.6
Method to estimate mutation efficiency Restriction enzyme digestionNlaIII(CATG)
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1021.pdf
Mutation efficiency in F0 embryos (%) 10
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 5%
Wild-type (WT) sequence AGTTTCTGGCCTTCTTCGCCATGTACACGGCCATCAGCATCACGTACAGgtgtgtgcatg
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 AGTTTCTGGCCTTCTTCGCAATGGACGCCAGGGACGACACGGCCATCAGGCTACGCGTACG Ggtgtgtgcatg
F0-2 AGTTTCTGGCCTTCTTCGCACACGGCCATCAGCATCACGTACA Ggtgtgtgcatg
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 AGTTTCTGGCCTTCTTCGCAATGGACGCCAGGGACGACACGGCCATCAGGCTACGCGTACG Ggtgtgtgcatg (-29bp,+42bp)
F0-2 AGTTTCTGGCCTTCTTCGCACACGGCCATCAGCATCACGTACA Ggtgtgtgcatg(-5bp)
Allele name and predicted product 1021a
1021b
AGTTTCTGGCCTTCTTCGCAATGGACGCCAGGGACGACACGGCCATCAGGCTACGCGTACG Ggtgtgtgcatg(splicing site)
AGTTTCTGGCCTTCTTCGCACACGGCCATCAGCATCACGTACAGgtgtgtgcatg(splicing site)
突变创建时间 2014-06-29
填表时间 2013/12/23
修改时间 2016/10/24
PI/负责人 (工作单位)
Genotype Information: