| ZKO number |
ZKO1161 |
| Recent progress of this line |
F2 generation has been raised
|
|
Gene name |
zgc:65894
|
|
Ensembl ID |
TUBB8P7 (1 of 3) ENSDARG00000016301
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGATCTAGAGCCTGGAACCA
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
exon3
|
|
Restriction enzyme (RE) |
n/a
|
|
PCR primer (forward) |
5’- TCTGGGAGGTGATCAGCGATG -3’
|
|
PCR primer (reverse) |
5’- CACCAAACTTGTTCCTGAAGATCC -3’
|
|
Length of expected PCR product (bp)
|
525
|
|
Length of expected RE digestion product (bp) |
|
|
Plasmids used for making Cas9 mRNA |
pGH-T7-zCAS9
|
| Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
| Primers?used?for?making?gRNA |
Pr-ZKO
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
600
|
|
Dosage of injected gRNA (ng/ul)
|
100
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
80
|
|
Method to estimate mutation efficiency |
PCR and single clone sequencing
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1161.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
|
16.7%
|
|
Wild-type (WT) sequence
|
TCCTTGTGGATCTAGAGCCTGGAACCATGGATTCA
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
TCCT---------------------------------AA----ATGGCTCA(22bp deletion, 2bp insertion)
|
|
F0-2
|
TCCTTGTGGATCTAGAGCC-------ATGGATTCA(7bp deletion)
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
TCCT---------------------------------AA----ATGGCTCA(-20bp, 1bp mutation)
|
|
F0-2
|
TCCTTGTGGATCTAGAGCC-------ATGGATTCA(-7bp)
TCTAGAGCCTGGATTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGACCT(30bp insertion, 7bp deletion)
|
|
Allele name and predicted product
|
1161a
1161b |
GTGGAAAGTATGTCCCTCGTGCCATCCTAAATGGACTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGACCTGACAACTTTGTTTTTG
|
| GTGGAAAGTATGTCCCTCGTGCCATCCTTGTGGATCTAGAGCCTGGATTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGTCCGCTCGGGTCCTTTTGGACAGATATTCAGACCTGACAACTTTGTTTTTG |
|
突变创建时间 |
|
|
填表时间 |
2013/12/20
|
|
修改时间 |
2016/8/9
|
|
PI/负责人 (工作单位) |
|
