| ZKO number |
ZKO107 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
fam134a
|
|
Ensembl ID |
ENSDARG00000063270
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
sense strand
GGGTCTGGAGGCCTGGTTCCcgg
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon1
|
|
Restriction enzyme (RE) |
Stu I（AGG/CCT）
|
|
PCR primer (forward) |
5’- tccatgacgagcctctcct -3’…………51.8
|
|
PCR primer (reverse) |
5’- accacggtaagagtagcgct -3’…………50.9
|
|
Length of expected PCR product (bp)
|
438
|
|
Length of expected RE digestion product (bp) |
120bp + 318bp
|
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
| Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
| Primers?used?for?making?gRNA |
Pr-ZKO 107
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
1000
|
|
Dosage of injected gRNA (ng/ul)
|
300
|
|
Survival rate of F0 embryo @ 1dpf (%)
|
63
|
|
Method to estimate mutation efficiency |
sequencing of PCR product
|
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO107.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
33
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
2/15
|
|
Wild-type (WT) sequence
|
GAGGCCTGGTTCCCGGCGCCCGGTGGCGCGGAGGAGGAACCGGAG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
GAGGCCTGGTTCCCGGCGCCCGG
GAGGCC------------------------GCCCGG (+0bp，-11bp）
|
|
F0-2
|
GAGGCCTGGTTCCCGGCGCCCGGTGGCGCGGAGGAGGAACCGGAGCT
GAGG----------------------------------------------------------------------------AACCGGAGCT （+0bp， -33bp）
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
GAGGCC------------------------GCCCGG (+0bp，-11bp）
|
|
F0-2
|
GAGG----------------------------------------------------------------------------AACCGGAGCT （+0bp， -33bp）
|
|
Allele name and predicted product
|
107a
107b |
GAGGCC------------------------GCCCGG Truncated protein （115aa）
|
| |
|
突变创建时间 |
|
|
填表时间 |
2013/12/17
|
|
修改时间 |
2016/12/29
|
|
PI/负责人 (工作单位) |
|
