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ZKO number ZKO1022
Recent progress of this line F2 individuals have been indentified
Gene name rab3da
Ensembl ID ENSDARG00000018107 .7
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGCCGACGCCATCTAGAGGAggg
Cas9 target site 2 sequence
Position in the gene Exon2
Restriction enzyme (RE) XbaI(TCTAGA)
PCR primer (forward) gtcctgaagggccacagcact
PCR primer (reverse) GCCGACTGTGCTGACGAATGC
Length of expected PCR product (bp) 278
Length of expected RE digestion product (bp) 178,100
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 1022
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 100
Survival rate of F0 embryo @ 1dpf (%) 75.6
Method to estimate mutation efficiency Restriction enzyme digestionxbaI
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO1022.pdf
Mutation efficiency in F0 embryos (%) 20
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 10
Wild-type (WT) sequence ttcactgtaaactacttacatatttactttttctcccctcctctAGATGGCGTCGGCCAACGATACCCGTCATCAGACACAGGTGCAGAAA
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 ttcactgtaaactacttacatatttactttgatatgatatgatatGATACCCGTCATCAGACACAGGTGCAGAAA(-31bp/+15 bp)
F0-2 tgtaaactacttacatatttactttttctccccgatatgatatgatatGCGTCGGCCAACGATACCCGTCATCAGACACAGGTGCAG(-11bp/+15 bp)
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 ttcactgtaaactacttacatatttactttgatatgatatgatatGATACCCGTCATCAGACACAGGTGCAGAAA(-31bp/+15 bp)
F0-2 tgtaaactacttacatatttactttttctccccgatatgatatgatatGCGTCGGCCAACGATACCCGTCATCAGACACAGGTGCAG(-11bp/+15 bp)
Allele name and predicted product 1022a
1022b
ttcactgtaaactacttacatatttactttgatatgatatgatatGATACCCGTCATCAGACACAGGTGCAGAAA(splicing site)
tgtaaactacttacatatttactttttctccccgatatgatatgatatGCGTCGGCCAACGATACCCGTCATCAGACACAGGTGCAG(splicing site)
突变创建时间 2014-02-19
填表时间 2013/12/23
修改时间 2016/10/24
PI/负责人 (工作单位)
Genotype Information: