China Zebrafish Resource Center (CZRC)

If you need ZKO Alleles,please contact luyuanpan@ihb.ac.cn

ZKO number		ZKO999
Recent progress of this line		F0 individuals have been indentified
Gene name		actr2a
Ensembl ID		ENSDARG00000052438
Zebrafish strain		AB
Cas9 target site 1 sequence		sense strand GGTCTGTGACAACGGGACAGggg
Cas9 target site 2 sequence
Position in the gene		exon1
Restriction enzyme (RE)		n/a
PCR primer (forward)		CTCACCCGTTTAACTCTTGTGG
PCR primer (reverse)		CTTCACTTCCTGTTCCGCTGC
Length of expected PCR product (bp)		388
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA		pXT7-hCAS9
Plasmids used for making gRNA		pMD19T-gRNA
Primers used for making gRNA		Pr-ZKO 999
Dosage of injected Cas9 mRNA (ng/ul)		35
Dosage of injected gRNA (ng/ul)		300
Survival rate of F0 embryo @ 1dpf (%)		20
Method to estimate mutation efficiency		sequencing of PCR product
Stage of the embryos collected for mutation detection		http://www.zfish.cn/upload/file/ZKOPDF/ZKO999.pdf
Mutation efficiency in F0 embryos (%)		37.5
Picture from F₀ embryos (gel picture or sequencing picture)		/upload/2014-09-02/635452689369541477.png 2014-09-02_153452
Mutation type		Indel
Germline transmission efficiency in adult F₀		100*3/8
Wild-type (WT) sequence		GGTCTGTGACAACGGGACAGGGG
Germline mutations in selected F0 (By screening F1 embryos)	F0-1
Germline mutations in selected F0 (By screening F1 embryos)	F0-2
Mosaicism in selected F0 (By screening F1 adults)	F0-1	GGTCTGTGACAAC-----AGGGG
Mosaicism in selected F0 (By screening F1 adults)	F0-2
Allele name and predicted product	999a 999b	GGTCTGTGACAAC-----AGGGG truncated protein（23aa）
Allele name and predicted product	999a 999b
突变创建时间
填表时间		2013/12/18
修改时间		2016/8/11
PI/负责人 (工作单位)

Genotype Information: