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ZKO number ZKO200
Recent progress of this line F2 individuals have been indentified
Gene name pmm2
Ensembl ID ENSDARG00000037654
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
GGCAGATGTTGTACACGTGGagg
Cas9 target site 2 sequence
Position in the gene exon1
Restriction enzyme (RE) BsrGI
PCR primer (forward) TCCGAACTTCCCCGCACTAT
PCR primer (reverse) ATTTCAGTCACTCCGCGTCA
Length of expected PCR product (bp) 582
Length of expected RE digestion product (bp) 173+409
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 200
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 90
Method to estimate mutation efficiency Restriction enzyme digestionBsrGI
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO200.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 45%
Wild-type (WT) sequence GGCCCATACATAGGGAAAACAGGTTATCCTTCTTATTTATGAAGTTGTTTGTCTGCTGTTGGCAGATGTTGTACACGTGGAGGTCGCTCACTCTGACGTACAGCTCTGGCGCTTCAATACACTTCCGACAAC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GGCCCATACATAGGGAAAACAGGTTATCCTTCTTATTTATGAAGTTGTTTGTCTGCTGTTGGCAGATGTTGTACGAGGTCGCTCACTCTGACGTACAGCTCTGGCGCTTCAATACACTTCCGACAAC -5bp
F0-2 GGCCCATACATAGGGAAAACAGGTTATCCTTCTTATTTATGAAGTTGTTTGTCTGCTGTTGGCAGGTGGAGGTCGCTCACTCTGACGTACAGCTCTGGCGCTTCAATACACTTCCGACAAC -11bp
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGCCCATACATAGGGAAAACAGGTTATCCTTCTTATTTATGAAGTTGTTTGTCTGCTGTTGGCAGATGTTGTACGAGGTCGCTCACTCTGACGTACAGCTCTGGCGCTTCAATACACTTCCGACAAC -5bp
F0-2 GGCCCATACATAGGGAAAACAGGTTATCCTTCTTATTTATGAAGTTGTTTGTCTGCTGTTGGCAGGTGGAGGTCGCTCACTCTGACGTACAGCTCTGGCGCTTCAATACACTTCCGACAAC -11bp
Allele name and predicted product 200a
200b
GGCCCATACATAGGGAAAACAGGTTATCCTTCTTATTTATGAAGTTGTTTGTCTGCTGTTGGCAGATGTTGTACGAGGTCGCTCACTCTGACGTACAGCTCTGGCGCTTCAATACACTTCCGACAAC
GGCCCATACATAGGGAAAACAGGTTATCCTTCTTATTTATGAAGTTGTTTGTCTGCTGTTGGCAGGTGGAGGTCGCTCACTCTGACGTACAGCTCTGGCGCTTCAATACACTTCCGACAAC
突变创建时间 2014-05-14
填表时间 2013/12/17
修改时间 2016/12/30
PI/负责人 (工作单位)
Genotype Information: