Dergi
/
Genetik
/
A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
JoVE Journal
Genetik
Bu içeriği görüntülemek için JoVE aboneliği gereklidir.  Oturum açın veya ücretsiz deneme sürümünü başlatın.
JoVE Journal Genetik
A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

DOI:
10.3791/57518-v

•

08:37 min

•

April 30, 2018

•

, , , ,
1Department of Pharmacology,University of Michigan

Bölümler

  • 00:04Başlık
  • 00:42sgRNA Target Selection and Homology-directed Repair Template Design
  • 02:21Prepare Injection Mix and Injection Protocol
  • 03:18Screen P0 Plates and Single mCherry(+) F1s
  • 04:03Single Worm PCR and Genotyping
  • 05:41Indentification and Sequence Verification of Edited Animals
  • 06:34Results: Rapid CRISPR/Cas9 Generated Point Mutations in C. elegans sod-1
  • 07:50Conclusion

Özet

Otomatik Çeviri

Otomatik Çeviri

Here, we present a method to engineer the genome of C. elegans using CRISPR-Cas9 ribonucleoproteins and homology dependent repair templates.

Etiketler

CRISPR/Cas9Genomic Point MutationC. ElegansHomology Directed RepairSsODNSgRNARestriction Endonuclease SiteNörobilimNeurodegenerative DiseaseEndogenous Regulatory ControlProtein OverexpressionPipelineRapidFacile

Article

Embed

ÇALMA LİSTESİNE EKLE

Usage Statistics

İlgili Videolar

Optogenetic Random Mutagenesis Using Histone-miniSOG in C. elegans

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format

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

Primordial Germ Cell Transplantation for CRISPR/Cas9-based Leapfrogging in Xenopus

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

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

CRISPR/Cas9 Gene Editing to Make Conditional Mutants of Human Malaria Parasite P. falciparum

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation

Read Article