NDLI: Integration of Double-Fluorescence Expression Vectors into Zebrafish Genome for the Selection of Site-Directed Knockout/Knockin

Content Provider	Springer Nature Link
Author	Wu, Yuping Zhang, Guangxian Xiong, Qian Luo, Fang Cui, Caimei Hu, Wei Yu, Yanhong Su, Jin Xu, Anlong Zhu, Zuoyan
Copyright Year	2006
Abstract	Production of zebrafish by modifying endogenous growth hormone (GH) gene through homologous recombination is described here. We first constructed the targeting vectors pGHT1.7k and pGHT2.8k, which were used for the knockout/knockin of the endogenous GH gene of zebrafish, and injected these two vectors into the embryos of zebrafish. Overall, the rate of targeted integration with the characteristic of germ line transmission in zebrafish was 1.7×10−6. In one experimental patch, the integrating efficiency of pGHT2.8k was higher than that of pGHT1.7k, but the lethal effect of pGHT2.8k was stronger than that of pGHT1.7k. The clones with the correct integration of target genes were identified by a simple screening procedure based on green fluorescent protein (GFP) and RFP dual selection, which corresponded to homologous recombination and random insertion, respectively. The potential homologous recombination zebrafish was further bred to produce a heterozygous F1 generation, selected based on the presence of GFP. The potential targeted integration of exogenous GH genes into a zebrafish genome at the P0 generation was further verified by polymerase chain reaction and Southern blot analysis. Approximately 2.5% of potential founder knockout and knockin zebrafish had the characteristic of germ line transmission. In this study, we developed an efficient method for producing the targeted gene modification in zebrafish for future studies on genetic modifications and gene functions using this model organism.
Starting Page	304
Ending Page	311
Page Count	8
File Format	PDF
ISSN	14362228
Journal	Marine Biotechnology
Volume Number	8
Issue Number	3
e-ISSN	14362236
Language	English
Publisher	Springer-Verlag
Publisher Date	2006-02-28
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	GH gene integration knockin knockout zebrafish Oceanography
Content Type	Text
Resource Type	Article
Subject	Aquatic Science Biotechnology

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Site-Directed Gene Integration in Transgenic Zebrafish Mediated by Cre Recombinase Using a Combination of Mutant Lox Sites

Efficient Transfection Strategy for the Spatiotemporal Control of Gene Expression in Zebrafish

Generation of Aberrant Transcripts of and Free DNA Ends in Zebrafish no tail Gene

Dicistronic Gene Expression in Developing Zebrafish

Baculovirus-Mediated Gene Expression in Zebrafish

RecA-Mediated, Targeted Mutagenesis in Zebrafish

Replacement of Buried Cysteine from Zebrafish Cu/Zn Superoxide Dismutase and Enhancement of Its Stability via Site-Directed Mutagenesis

Expression of Masu Salmon Δ5-Desaturase-Like Gene Elevated EPA and DHA Biosynthesis in Zebrafish

Learning from Small Fry: The Zebrafish as a Genetic Model Organism for Aquaculture Fish Species

Integration of Double-Fluorescence Expression Vectors into Zebrafish Genome for the Selection of Site-Directed Knockout/Knockin

Similar Documents

Site-Directed Gene Integration in Transgenic Zebrafish Mediated by Cre Recombinase Using a Combination of Mutant Lox Sites

Efficient Transfection Strategy for the Spatiotemporal Control of Gene Expression in Zebrafish

Generation of Aberrant Transcripts of and Free DNA Ends in Zebrafish no tail Gene

Dicistronic Gene Expression in Developing Zebrafish

Baculovirus-Mediated Gene Expression in Zebrafish

RecA-Mediated, Targeted Mutagenesis in Zebrafish

Replacement of Buried Cysteine from Zebrafish Cu/Zn Superoxide Dismutase and Enhancement of Its Stability via Site-Directed Mutagenesis

Expression of Masu Salmon Δ5-Desaturase-Like Gene Elevated EPA and DHA Biosynthesis in Zebrafish

Learning from Small Fry: The Zebrafish as a Genetic Model Organism for Aquaculture Fish Species

Integration of Double-Fluorescence Expression Vectors into Zebrafish Genome for the Selection of Site-Directed Knockout/Knockin