If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO201
Recent progress of this line F1 individuals have been identified
Gene name C1H7orf26
Ensembl ID ENSDARG00000037652
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGACTAGAGAAGAGCACAGtgg
Cas9 target site 2 sequence
Position in the gene exon2
Restriction enzyme (RE) BspQI(GCTCTTC)
PCR primer (forward) TTCCTCCTCCACAGCAGAGA
PCR primer (reverse) TCCCTTTAAAGCGTACCTTCCT
Length of expected PCR product (bp) 320
Length of expected RE digestion product (bp) 143+177
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 201
Dosage of injected Cas9 mRNA (ng/ul) 600
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 95
Method to estimate mutation efficiency Restriction enzyme digestionBspQI
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO201.pdf
Mutation efficiency in F0 embryos (%) 30
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 25
Wild-type (WT) sequence TGCAGAGCTCCGCTGGACCCCCGGCTGGACTAGAGAAGAGCACAGTGGAGCTGGTGGAAGAGTTCATCTTTCA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 22%(4/18) TGCAGAGCTCCGCTGGACCCCCGGCTGGACxxxxxxxxxxxxxAGTGGAGCTGGTGGAAGAGTTCATCTTTCA (-13bp)
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 27%(9/33) TGCAGAGCTCCGCTGGACCCCCGGCTGGACxxxxxxxxxxxxxAGTGGAGCTGGTGGAAGAGTTCATCTTTCA (-13bp)
F0-2
Allele name and predicted product 201a
201b
TCTGCAGAGCTCCGCTGGACCCCCGGCTGGACxxxxxxxxxxxxxAGTGGAGCTGGTGGAAGAGTTCATCTTT;Truncated protein(66aa)
突变创建时间 2013-10-09
填表时间 2013/12/18
修改时间 2016/12/30
PI/负责人 (工作单位)
Genotype Information: