If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1035
Recent progress of this line F1 individuals have been identified
Gene name zgc:100913
Ensembl ID ENSDARG00000052386
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
gagccgatcaccagcgctg
Cas9 target site 2 sequence
Position in the gene Exon 2
Restriction enzyme (RE) BstX1
PCR primer (forward) CGTTTCGCATTGATGAGCTTGA
PCR primer (reverse) AAACCAATGCGAGTCTTGATGC
Length of expected PCR product (bp) 460
Length of expected RE digestion product (bp) 200/280
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1035
Dosage of injected Cas9 mRNA (ng/ul) 320
Dosage of injected gRNA (ng/ul) 40
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1035.pdf
Mutation efficiency in F0 embryos (%) 80
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 CTGAAAGTCATTTTTGCAACAAAAAACAACATGGTGGAC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1
F0-2 CTGAAAGTCATCTTTACAAC----AACAACATGGTGGAC(-13bp,+9bp)
Allele name and predicted product 1035a
1035b
CTGAAAGTCATCTTTACAAC----AACAACATGGTGGAC (out frame mutation)
突变创建时间
填表时间 2014/1/3
修改时间 2016/11/11
PI/负责人 (工作单位)
Genotype Information: