If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1163
Recent progress of this line F2 individuals have been indentified
Gene name c3b
Ensembl ID ENSDARG00000087359
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GAGCCATCAGAACATACCTGagg
Cas9 target site 2 sequence
Position in the gene junction of intron 1 and exon2
Restriction enzyme (RE) n/a
PCR primer (forward) 5’- CGTGCCTGGTTACGGACAA -3’
PCR primer (reverse) 5’- GTGACGGAGGGAAAGTGAG -3’
Length of expected PCR product (bp) 545
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1163
Dosage of injected Cas9 mRNA (ng/ul) 650
Dosage of injected gRNA (ng/ul) 80
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency PCR and single clone sequencing
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1163.pdf
Mutation efficiency in F0 embryos (%) 20
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 50%
Wild-type (WT) sequence GTTCTCCTCAGGTATGTTCTGATGGCTCCTAATCTG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GTTCT---------------GATAGCTCCTAATCTG(-15bp)
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GTTCT---------------GATAGCTCCTAATCTG(-15bp)
F0-2
Allele name and predicted product 1163a
1163b
change splicing site in intron 1/exon 2
突变创建时间 2014-12-03
填表时间 2013/12/20
修改时间 2016/8/9
PI/负责人 (工作单位)
Genotype Information: