| 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) |
|