If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO339
Recent progress of this line F1 individuals have been identified
Gene name ufsp2
Ensembl ID ENSDARG00000056369
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGCTGTGGCACCAGGAACGagg
Cas9 target site 2 sequence
Position in the gene Exon 7
Restriction enzyme (RE) Sty D4I(|CCNGG)
PCR primer (forward) AGTGTCTTTACAGGTCTGGTGGA
PCR primer (reverse) GGACTGTTGTGGTAAGTTTATTTTG
Length of expected PCR product (bp) 490
Length of expected RE digestion product (bp) 89+401
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 339
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 30
Survival rate of F0 embryo @ 1dpf (%) 95
Method to estimate mutation efficiency Restriction enzyme digestionRE digestion (Sty D4I)
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO339.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 28%(4/14)
Wild-type (WT) sequence GGCTGTGGCACCAGGAACGAGGAGCCCTTCC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGCTGTGGCACCAGCTCCAACGAGGAGCCCTTCC(+4bp,-1bp)
F0-2 GGCTGTGGCACCAGGA-------------CC(-13bp)测序鉴定
Allele name and predicted product 339a
339b
GGCTGTGGCACCAGCTCCAACGAGGAGCCCTTCC; Truncated protein(402aa)
GGCTGTGGCACCAGGA-------------CC; Truncated protein(139aa)
突变创建时间 2014-07-02
填表时间 2013/12/18
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information: