If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO331
Recent progress of this line F2 generation has been raised
Gene name casp3a
Ensembl ID ENSDARG00000017905
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGCATCTGCATCACAGCCCATGCagg
Cas9 target site 2 sequence
Position in the gene Exon 3
Restriction enzyme (RE) n/a
PCR primer (forward) ATAAGTAGAAAACTCACCCACAG
PCR primer (reverse) ACACAAGACAACTCACACCG
Length of expected PCR product (bp) 297
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO
Dosage of injected Cas9 mRNA (ng/ul) 400
Dosage of injected gRNA (ng/ul) 50
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/ZKO331.pdf
Mutation efficiency in F0 embryos (%) 40
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 50%(2/4)
Wild-type (WT) sequence AGGATGGGGCATCTGCATCACAGCCCATGCAGGTTGATGCCAAGCC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 AGGATGGGGCATCTGCATCACAGCCCA----GGTTGATGCCAAGCC (-4)
F0-2 AGGATGGGGCATCTGCATCACAGCCCATGTATATGAGCCAAGCC(-9,+7)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 AGGATGGGGCATCTGCATCACAGCCCA----GGTTGATGCCAAGCC (-4)
F0-2
Allele name and predicted product 331a
331b
AGGATGGGGCATCTGCATCACAGCCCA----GGTTGATGCCAAGCC (40aa)
突变创建时间 2013-05-17
填表时间 2013/12/18
修改时间 2016/10/14
PI/负责人 (工作单位)
Genotype Information: