| ZKO number |
ZKO352 |
| Recent progress of this line |
F2 individuals have been indentified
|
|
Gene name |
klf3
|
|
Ensembl ID |
ENSDARG00000015495
|
|
Zebrafish strain |
AB
|
|
Cas9 target site 1 sequence |
antisense strand
5’- GGGTGCGGCTCAGATTTGAT tgg -3’
|
|
Cas9 target site 2 sequence |
|
|
Position in the gene |
Exon 5
|
|
Restriction enzyme (RE) |
T7E1
|
|
PCR primer (forward) |
5’- TGTCTCTTGTCTTCAGTCTCTGT -3’
|
|
PCR primer (reverse) |
5’- AGACAGCAGCATTTGTGTAAT -3’
|
|
Length of expected PCR product (bp)
|
411
|
|
Length of expected RE digestion product (bp) |
~88+~322
|
|
Plasmids used for making Cas9 mRNA |
pSP6-2sNLS-spCas9
|
| Plasmids?used?for?making?gRNA |
pT7-gRNA-ZKO352
|
| Primers?used?for?making?gRNA |
Pr-ZKO 352
|
|
Dosage of injected Cas9 mRNA (ng/ul)
|
450
|
|
Dosage of injected gRNA (ng/ul)
|
80
|
|
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/ZKO352.pdf
|
|
Mutation efficiency in F0 embryos (%)
|
10
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
|
Mutation type
|
Indel
|
|
Germline transmission efficiency in adult F0
|
36% (5/14: Number of positive F0 / Total number of F0 screened)
|
|
Wild-type (WT) sequence
|
GAGCTTCTCAAACCAATCAAATCTGAGCCGCACCCTGAACATACTCAG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
GAGCTTCTCAAACCAATCxxxxxTGAGCCGCACCCTGAACATACTCAGG(- 5 bp)
|
|
F0-2
|
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
GAGCTTCTCAAACCAATCxxxxxTGAGCCGCACCCTGAACATACTCAGG(- 5 bp) (- 5 bp)
|
|
F0-2
|
|
|
Allele name and predicted product
|
352a
352b |
GAGCTTCTCAAACCAATCxxxxxTGAGCCGCACCCTGAACATACTCAGG(- 5 bp); Truncated protein (194 aa)
|
| |
|
突变创建时间 |
2014-10-08
|
|
填表时间 |
2013/12/24
|
|
修改时间 |
2016/8/8
|
|
PI/负责人 (工作单位) |
|
