ZKO number ZKO16
Recent progress of this line F1 individuals have been identified
Gene name rasa3
Ensembl ID ENSDARG00000063371
Zebrafish strain AB
Cas9 target site 1 sequence sense strand
5’- GGAGAATCTGGACACAAACA ggg -3’
Cas9 target site 2 sequence
Position in the gene Exon 13
Restriction enzyme (RE) T7EI
PCR primer (forward) 5’- TGTGCATACACCCTATTGGTTAC -3’
PCR primer (reverse) 5’- CTCATTTCATTTGTCTTTTGTACTG -3’
Length of expected PCR product (bp) 500
Length of expected RE digestion product (bp) 330,270
Plasmids used for making Cas9 mRNA pXT7-hCAS9
Plasmids used for making gRNA pMD19T-gRNA
Primers used for making gRNA Pr-ZKO 16
Dosage of injected Cas9 mRNA (ng/ul) 300
Dosage of injected gRNA (ng/ul) 60
Survival rate of F0 embryo @ 1dpf (%) 80
Method to estimate mutation efficiency T7EI detection
Stage of the embryos collected for mutation detection http://www.zfish.cn/upload/file/ZKOPDF/ZKO16.pdf
Mutation efficiency in F0 embryos (%) 60
Picture from F0 embryos
  (gel picture or sequencing picture)
Mutation type Indel
Germline transmission efficiency in adult F0 20%
Wild-type (WT) sequence tcggagaatctggacaca a acagggaaaacctgcg
Germline mutations in selected F0
 (By screening F1 embryos)
F0-1
F0-2
Mosaicism in selected F0
 (By screening F1 adults)
F0-1 tcggagaatctggacacaa- - - -ggaaaacctgcg (-4 bp)
F0-2 tcggagaatctggacacaCaGacagggaaaacctgcg (+2 bp)
Allele name and predicted product 16a
16b
tcggagaatctggacacaa- - - -ggaaaacctgcg; Truncated protein (437 aa)
tcggagaatctggacacaCaGacagggaaaacctgcg; Truncated protein (439 aa)
突变创建时间
填表时间 2013/12/24
修改时间 2016/8/8
PI/负责人 (工作单位)
Genotye Information: