国家水生生物种质资源库国家斑马鱼资源中心 (China Zebrafish Resource Center)

ZKO number		ZKO1194
Recent progress of this line		F1 individuals have been identified
Gene name		epsin 3b
Ensembl ID		ENSDARG00000004633
Zebrafish strain		AB
Cas9 target site 1 sequence		sense strand GGCGACCTCTAATGACCCCTggg
Cas9 target site 2 sequence
Position in the gene		exon1
Restriction enzyme (RE)		n/a
PCR primer (forward)		aggacagctgagtcctcaca
PCR primer (reverse)		GATAATCCAGCAGCGTCA
Length of expected PCR product (bp)		453
Length of expected RE digestion product (bp)
Plasmids used for making Cas9 mRNA		pGH-T7-zCAS9
Plasmids?used?for?making?gRNA		pMD19T-gRNA
Primers?used?for?making?gRNA		Pr-ZKO 1194
Dosage of injected Cas9 mRNA (ng/ul)		300
Dosage of injected gRNA (ng/ul)		100
Survival rate of F0 embryo @ 1dpf (%)		89
Method to estimate mutation efficiency		sequencing of PCR product
Stage of the embryos collected for mutation detection		3dpf
Mutation efficiency in F0 embryos (%)		60
Picture?from?F₀?embryos ? (gel?picture?or?sequencing?picture)		/upload/2015-09-29/635791228970802505.png undefined QQ图片20150929110520
Mutation type		deletion
Germline transmission efficiency in adult F₀		50%（4/8）
Wild-type (WT) sequence		AGGCGACCTCTAATGACCCCTGGGGCCCGTCCAGCTC
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	AGGCGACCTCTAATG - - - - - - - GGGCCCGTCCAGCTC
Mosaicism in selected F0 (By screening F1 adults)	F0-2
Allele name and predicted product	1194a 1194b	AGGCGACCTCTAATG - - - - - - - GGGCCCGTCCAGCTC Truncated protein（61aa）
Allele name and predicted product	1194a 1194b
突变创建时间		2014-10-15
填表时间		2013/12/16
修改时间		2015/10/8
PI/负责人 (工作单位)		殷战(中科院水生所)

Genotype Information: