If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO180
Recent progress of this line F2 individuals have been indentified
Gene name vap
Ensembl ID ENSDARG00000093053
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
5'-GAGAGCATAGTGGCTCAATC cgg-3'
Cas9 target site 2 sequence
Position in the gene exon 2
Restriction enzyme (RE) T7E1
PCR primer (forward) 5’- GCGTACTTTACAAGTGTAATTCTGG -3’
PCR primer (reverse) 5’- ACAACAGTTCTCCCAGTGCC -3’
Length of expected PCR product (bp) 468
Length of expected RE digestion product (bp) ~231 +~236
Plasmids used for making Cas9 mRNA pSP6-2sNLS-spCas9
Plasmids used for making gRNA pT7-gRNA-ZKO180
Primers used for making gRNA Pr-ZKO 180
Dosage of injected Cas9 mRNA (ng/ul) 450
Dosage of injected gRNA (ng/ul) 60
Survival rate of F0 embryo @ 1dpf (%) 60
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO180.pdf
Mutation efficiency in F0 embryos (%) 30
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 67% (4/6: Number of positive F0 / Total number of F0 screened)
Wild-type (WT) sequence GAGAGCATAGTGGCTCAATC cgg
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GAGAGCATAGTGGCTCxxTC cgg(-2 bp)
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GAGAGCATAGTGGCTCxxTC cgg(-2 bp)
F0-2
Allele name and predicted product 180a
180b
GAGAGCATAGTGGCTCxxTC; Truncated protein (47 aa)
突变创建时间 2014-09-30
填表时间 2013/12/24
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information: