If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO329
Recent progress of this line F1 progeny has been raised
Gene name fat1
Ensembl ID ENSDARG00000017591
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGTTGCACTTCACGATCTAGagg
Cas9 target site 2 sequence
Position in the gene exon2
Restriction enzyme (RE) n/a
PCR primer (forward) 5’- CTGTGGACTGTTGTCATTAGC -3’
PCR primer (reverse) 5’- CACTAGAGGATGTAAACGACAAC -3’
Length of expected PCR product (bp) 327
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pSP6-2sNLS-spCas9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 329
Dosage of injected Cas9 mRNA (ng/ul) 600
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO329.pdf
Mutation efficiency in F0 embryos (%) 8
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 1/12
Wild-type (WT) sequence GGTTGCACTTCACGATCTAGAGG
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GGTTGCACTTCACGA------GG △6
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1
F0-2
Allele name and predicted product 329a
329b
GGTTGCACTTCACGA------GG truncated protein (4609aa)
突变创建时间 2016-04-20
填表时间 2013/12/18
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information: