If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO210
Recent progress of this line F2 generation has been raised
Gene name rnf175
Ensembl ID ENSDARG00000037647
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGCTACACGTGAAGCACAGagg
Cas9 target site 2 sequence
Position in the gene exon4
Restriction enzyme (RE) n/a
PCR primer (forward) TGCTTCAGTATATCTTCCTTCAT
PCR primer (reverse) TGTGTGTAAGCCAACAGATCA
Length of expected PCR product (bp) 381
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 210
Dosage of injected Cas9 mRNA (ng/ul) 400
Dosage of injected gRNA (ng/ul) 60
Survival rate of F0 embryo @ 1dpf (%) 75
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO210.pdf
Mutation efficiency in F0 embryos (%) 35
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0
Wild-type (WT) sequence AGGCTACACGTGAAGCACAGAGGTCATGA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 CGGCTACACGT--------GAGGTCATGA
F0-2 AGGCTACACGTGA----TGGCCTCAGAGGTCATGA
Allele name and predicted product 210a
210b
CGGCTACACGT--------GAGGTCATGA truncated protein (18aa)
AGGCTACACGTGA----TGGCCTCAGAGGTCATGA truncated protein (30aa)
突变创建时间 2013-07-08
填表时间 2013/12/18
修改时间 2016/10/14
PI/负责人 (工作单位)
Genotype Information: