If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO100
Recent progress of this line F2 individuals have been indentified
Gene name nrp2a
Ensembl ID ENSDARG00000031622
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GCCACAGTATCGTCCGATCAAggg
Cas9 target site 2 sequence
Position in the gene Exon2
Restriction enzyme (RE) Bsl I (CCNNNNN/NNGG)
PCR primer (forward) 5’- ctcgggatcataggagcagt -3’…………50.7
PCR primer (reverse) 5’- tcatgggaccttcaaatgtg -3’…………50.4
Length of expected PCR product (bp) 437
Length of expected RE digestion product (bp) 214bp +223 bp
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 100
Dosage of injected Cas9 mRNA (ng/ul) 1000
Dosage of injected gRNA (ng/ul) 300
Survival rate of F0 embryo @ 1dpf (%) 75.6
Method to estimate mutation efficiency Restriction enzyme digestion60
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO100.pdf
Mutation efficiency in F0 embryos (%) 50
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 50% (5/10)
Wild-type (WT) sequence tccaacatttccttgtgttcagtGGGACCCTTGATCGGACGATACTGTGGCTCGAAGATCCCTCCAAAGATCCAGTCCTCC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 tccaacatttccttgtgttcagtGGGACCCTTTGGAGGGACCCTTGAAGATCCCTCCAAAGATCCAGTCCTCC (+13bp, -21 bp) sense
F0-2 tccaacatttccttgtgttcagtGGGACCCTT--------------------------------------------GATCCCTCCAAAGATCCAGTCCTCC (+0bp, -24bp) sense
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 tccaacatttccttgtgttcagtGGGACCCTTTGGAGGGACCCTTGAAGATCCCTCCAAAGATCCAGTCCTCC (+13bp, -21 bp) sense
F0-2 tccaacatttccttgtgttcagtGGGACCCTT--------------------------------------------GATCCCTCCAAAGATCCAGTCCTCC (+0bp, -24bp) sense
Allele name and predicted product 100a
100b
tccaacatttccttgtgttcagtGGGACCCTTTGGAGGGACCCTTGAAGATCCCTCCAAAGATCCAGTCCTCC truncnted protein (108aa)
cttgtgttcagtGGGACCCTT--------------------------------------------GATCCCTCCAAAGATCCA functional domain lose protein
突变创建时间
填表时间 2013/12/16
修改时间 2016/12/29
PI/负责人 (工作单位)
Genotype Information: