If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO995
Recent progress of this line F2 individuals have been indentified
Gene name ndufaf5
Ensembl ID ENSDARG00000061629
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGATTGGGCTTCATCTTTAC tgg
Cas9 target site 2 sequence
Position in the gene exon 1
Restriction enzyme (RE) n/a
PCR primer (forward) CTGACTCCGATCCGTGTTCC
PCR primer (reverse) AGCATAGTGACCTTGCGACA
Length of expected PCR product (bp) 489
Length of expected RE digestion product (bp) n/a
Plasmids used for making Cas9 mRNA pGH-T7-zCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 995
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 100
Survival rate of F0 embryo @ 1dpf (%) 85
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO995.pdf
Mutation efficiency in F0 embryos (%) 50
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type deletion
Germline transmission efficiency in adult F0 14.3%(2/14)
Wild-type (WT) sequence TCTTTACTGGACAGCAGCAAAT
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 25%(2/8) TCT--ACTGGACAGCAGCAAAT(-2bp)
F0-2 33.3%(2/6) TCT----TGGACAGCAGCAAAT(-4bp)
Allele name and predicted product 995a
995b
TCT--ACTGGACAGCAGCAAAT(-2bp),Truncated protein (88aa)
TCT----TGGACAGCAGCAAAT(-4bp),Truncated protein (75aa)
突变创建时间
填表时间 2013/12/18
修改时间 2016/8/11
PI/负责人 (工作单位)
Genotype Information: