| ZKO number |
ZKO1065 |
| Recent progress of this line |
F1 individuals have been identified
|
|
Gene name |
null
|
|
Ensembl ID |
ENSDARG00000089742
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
antisense strand
TTTAAAGCCAAACCTGTCCC
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 1
|
|
Restriction enzyme (RE) |
Dra I (TTT/AAA)
|
|
PCR primer (forward) |
AATGAAGAGCGAAGAAGACT
|
|
PCR primer (reverse) |
TTTGTGCGATGTGATGTTAG
|
|
Length of expected PCR product (bp)
|
525
|
|
Length of expected RE digestion product (bp) |
177+348
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids used for making gRNA |
pMD19T-gRNA
|
| Primers used for making gRNA |
Pr-ZKO 1065
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
|
Dosage of injected gRNA (ng/ul)
|
50
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
91.6
|
|
Method to estimate mutation efficiency |
Restriction enzyme digestionRE digestion (BstYI)
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO1065.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
|
50% (1/2)
|
|
Wild-type (WT) sequence
|
agaggaagaggagaggaggaagtgcccgtttaaagccaaacctgtcccaaaggcggtgaa
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
60.7% (m/n: Number of F1 carrier / Total number of F1 screened)
CGCCCAACGTGGCTTCCTTCACCGCCTTTGGGACAGGTT-----------GGGCACTTCCTCCTCTCCTCTTCCTCTTTG (+0 bp, -11 bp)
|
|
F0-2
|
68.75% (m/n: Number of F1 carrier / Total number of F1 screened)
CGCCCAACGTGGCTTCCTTCACCGCCTTTGGGACAGGTTTGGCTTT---CGGGCACTTCCTCCTCTCCTCTTCCTCTTTG (+0 bp, -3 bp)
|
|
Allele name and predicted product
|
1065a
1065b |
daklreqtlqrnkeeeerrkcptcpkggegshvgraterggtlqgn* Truncated protein (207 aa)
|
| daklreqtlqrnkeeeerrkcp-kakpvpkavkeatlgerqkeeelyraikmqmraremlh; Truncated protein (609 aa) |
|
突变创建时间 |
|
|
填表时间 |
2014/1/4
|
|
修改时间 |
2016/11/11
|
|
PI/负责人 (工作单位) |
|