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ZKO number ZKO12
Recent progress of this line F2 individuals have been indentified
Gene name grk1a
Ensembl ID ENSDARG00000058803
Zebrafish strain AB
Cas9 target site 1 sequence antisense strand
GGCGGAGATGTAGGCAGAGT tgg
Cas9 target site 2 sequence
Position in the gene Exon 1
Restriction enzyme (RE) T7E1
PCR primer (forward) TTATCGTCCACTTGTCCATCA
PCR primer (reverse) CGTATTCCTCTACGTCCCTCC
Length of expected PCR product (bp) 466
Length of expected RE digestion product (bp) 231+235
Plasmids used for making Cas9 mRNA pSP6-2sNLS-spCas9
Plasmids?used?for?making?gRNA pT7-gRNA-ZKOpT7-gRNA-ZKO12
Primers?used?for?making?gRNA Pr-ZKO 12
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 50
Survival rate of F0 embryo @ 1dpf (%) 81.2
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO12.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 66.7% (10/15: Number of positive F0 / Total number of F0 screened)
Wild-type (WT) sequence GGCGGCCTCACCACGGTGGTG-GCCAACTCTGCC
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1 GGCGG------------------------------CTGCC(, -30 bp)
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 GGCGG------------------------------CTGCC(, -30 bp)
F0-2
Allele name and predicted product 12a
12b
GGCGG------------------------------CTGCC(, -30 bp) Truncated protein (10aa)
突变创建时间 2014-03-12
填表时间 2013/12/24
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotype Information:
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