ZKO number |
ZKO629 |
Recent progress of this line |
F2 generation has been raised
|
Gene name |
htf1fb
|
Ensembl ID |
ENSDARG00000053580
|
Zebrafish strain |
AB
|
Cas9 target site 1 sequence |
sense strand
GGACGGGAAGAGGGAACCCAtgg
|
Cas9 target site 2 sequence |
|
Position in the gene |
Exon 1
|
Restriction enzyme (RE) |
StyI (C/CWWGG)
|
PCR primer (forward) |
TTCCATTCTGGTGTCCCTTC
|
PCR primer (reverse) |
CCGCTGCCTTGTAGATCTTG
|
Length of expected PCR product (bp)
|
197
|
Length of expected RE digestion product (bp) |
78
|
Plasmids used for making Cas9 mRNA |
pXT7-hCAS9
|
Plasmids?used?for?making?gRNA |
pMD19T-gRNA
|
Primers?used?for?making?gRNA |
Pr-ZKO
|
Dosage of injected Cas9 mRNA (ng/ul)
|
300
|
Dosage of injected gRNA (ng/ul)
|
30
|
Survival rate of F0 embryo @ 1dpf (%)
|
31
|
Method to estimate mutation efficiency |
Restriction enzyme digestionstyI
|
Stage of the embryos collected for mutation detection |
http://www.zfish.cn/upload/file/ZKOPDF/ZKO629.pdf
|
Mutation efficiency in F0 embryos (%)
|
50
|
Picture?from?F0?embryos
? (gel?picture?or?sequencing?picture) |
|
Mutation type
|
Indel
|
Germline transmission efficiency in adult F0
|
27
|
Wild-type (WT) sequence
|
AAGAGGGAACCCATGGACTGTTTG
|
Germline mutations in selected F0
(By screening F1 embryos) |
F0-1 |
AAGAGGG----------ACTGTTTG (-9bp)
AAGAGGGAACCCATGTG-ACTGTTTG(+2 bp,-1bp)
|
F0-2
|
AAGAGGGAACTGTTGGACTGTTTG(change 3bp)
AAGAGG
|
Mosaicism in selected F0
(By screening F1 adults) |
F0-1
|
AAGAGGG----------ACTGTTTG (-9bp)
|
F0-2
|
AAGAGGGA----ACTGTTTG (-8bp)
|
Allele name and predicted product
|
629a 629b |
ZKO629a AAGAGGG------ACTGTTTG (delete 3aa)
|
ZKO629a AAGAGGGA----ACTGTTTG Truncated protein (174aa) |
突变创建时间 |
2013-05-01
|
填表时间 |
2013/12/23
|
修改时间 |
2016/10/24
|
PI/负责人 (工作单位) |
|