If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn
ZKO number ZKO336
Recent progress of this line F1 individuals have been identified
Gene name slc25a4
Ensembl ID ENSDARG00000027355
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGATTTCTTGGCCGGTGGTGtgg
Cas9 target site 2 sequence
Position in the gene Exon 1
Restriction enzyme (RE) n/a
PCR primer (forward) ATCAATCACCCGCTCCTCT
PCR primer (reverse) CCTTTATTCGCCTACTCTTCT
Length of expected PCR product (bp) 429
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO
Dosage of injected Cas9 mRNA (ng/ul) 400
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency sequencing of PCR product
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO336.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 33%(4/12)
Wild-type (WT) sequence ATCAGCTTCTTGAAGGATTTCTTGGCCGGTGGTGTGGCCGCTGCCATTTCCAAGACAGCA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 TTGAAGGATTTCTTGGCCGGT-----------------TTCCAAGACAGCAG (-17bp)
F0-2 GAGATAGGACATCATGTCTGA--------------TC-------A-------------------------G-------C—GACAGCAGTTGC(-60,+5)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 TTGAAGGATTTCTTGGCCGGT-----------------TTCCAAGACAGCAG (-17bp)
F0-2 GAGATAGGACATCATGTCTGA--------------TC-------A-------------------------G-------C—GACAGCAGTTGC(-60,+5)
Allele name and predicted product 336a
336b
TTGAAGGATTTCTTGGCCGGT-----------------TTCCAAGACAGCAG ( 23aa)
GAGATAGGACATCATGTCTGA--------------TC-------A-------------------------G-------C—GACAGCAGTTGC(40aa)
突变创建时间 2013-05-27
填表时间 2013/12/18
修改时间 2016/10/14
PI/负责人 (工作单位)
Genotype Information: