'Dergi'
/
Genetik
/
Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
JoVE Journal
Genetik
This content is Free Access.
JoVE Journal Genetik
Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

DOI:
10.3791/56969-v

•

11:27 min

•

August 28, 2018

•

, ,
1Department of Biochemistry,Purdue University

'Bölümler'

  • 00:05'Başlık'
  • 01:09Preparation of CRISPR-reagents for Embryo Microinjection
  • 04:03Microinjection of CRISPR-components into Zebrafish Embryos
  • 06:12Analysis of Efficiency of Indel Formation Using an HMA
  • 08:45Identification and Propagation of Knock-out Lines
  • 09:45Results: Zebrafish Embryos Exhibit a Pigment Defect when Injected with an sgRNA Targeting Tyrosinase at the One Cell Stage
  • 10:32Conclusion

Özet

'Otomatik Çeviri'

'Otomatik Çeviri'

Targeted genome editing in the model system Danio rerio (zebrafish) has been greatly facilitated by the emergence of CRISPR-based approaches. Herein, we describe a streamlined, robust protocol for generation and identification of CRISPR-derived nonsense alleles that incorporates the heteroduplex mobility assay and identification of mutations using next-generation sequencing.

'Etiketler'

CRISPR/Cas9Gene KnockoutDanio RerioZebrafishTargeted Genome EditingIn Vitro TranscriptionSgRNAAmmonium Acetate PrecipitationEthanol PrecipitationRNA Purification

Article

Embed

'ÇALMA LİSTESİNE EKLE'

Usage Statistics

'İlgili Videolar'

Rearing and Double-stranded RNA-mediated Gene Knockdown in the Hide Beetle, Dermestes maculatus

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells

Production and Purification of Baculovirus for Gene Therapy Application

Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9

Lentiviral Mediated Production of Transgenic Mice: A Simple and Highly Efficient Method for Direct Study of Founders

High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)

Maintaining Biological Cultures and Measuring Gene Expression in Aphis nerii: A Non-model System for Plant-insect Interactions

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System

Read Article