If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO1081
Recent progress of this line F1 individuals have been identified
Gene name rfx1b
Ensembl ID ENSDARG00000075904
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGCCTGTGCTCTGATGATGGagg
Cas9 target site 2 sequence
Position in the gene Exon 1
Restriction enzyme (RE) n/a
PCR primer (forward) TAAACCTGTGTGCCTTCG
PCR primer (reverse) TGTAGTGTGTAGTGTGCTCACC
Length of expected PCR product (bp) 409
Length of expected RE digestion product (bp) n/a
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1081
Dosage of injected Cas9 mRNA (ng/ul) 100
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 65
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1081.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 67%
Wild-type (WT) sequence GGATCGGCTTCCTCCATCATCAGAGCACAGGCCACGCCC
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 GGATCGGCTTCCTC-GGCTTCAGAGCACAGGCCACGCCC
Allele name and predicted product 1081a
1081b
GGATCGGCTTCCTC-GGCTTCAGAGCACAGGCCACGCCC Truncated protein (95aa)
突变创建时间 2013-12-10
填表时间 2013/12/20
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information: