If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO370
Recent progress of this line F2 individuals have been indentified
Gene name kit receptor b
Ensembl ID ENSDARG00000056133
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGGGGGCATCAGGAGTGATAAC
Cas9 target site 2 sequence
Position in the gene Exon2
Restriction enzyme (RE) AgeI
PCR primer (forward) GATAAAGCCCTTCTCATCTTCC
PCR primer (reverse) CTGATTCAAACTCACCTTTCAC
Length of expected PCR product (bp) 406
Length of expected RE digestion product (bp) 274bp+132bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 370
Dosage of injected Cas9 mRNA (ng/ul) 150
Dosage of injected gRNA (ng/ul) 150
Survival rate of F0 embryo @ 1dpf (%) 92
Method to estimate mutation efficiency Restriction enzyme digestionAgeI
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO370.pdf
Mutation efficiency in F0 embryos (%) 15
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 33.3
Wild-type (WT) sequence TTCCAGGATGGTCTAAACCGGTTATCACTCCTGATGCCCCCCATCTGGTGATTCTTAAAGGT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 TTCCAGGATGGTCTAA - - - - - - - - - - -CTCCTGATGCCCCCCATCTGGTGATTCTTAAAGGT
F0-2 TTCCAGGATGGTCTAAACCG - - - - - - - - - - - - - - - CCCCCCATCTGGTGATTCTTAAAGGT
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TTCCAGGATGGTCTAA - - - - - - - - - - -CTCCTGATGCCCCCCATCTGGTGATTCTTAAAGGT
F0-2 TTCCAGGATGGTCTAAACCG - - - - - - - - - - - - - - - CCCCCCATCTGGTGATTCTTAAAGGT
Allele name and predicted product 370a
370b
(-11,+0)
(-16,+0)
突变创建时间 2014-03-19
填表时间 2013/12/19
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information: