PUBLICATION
A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafish
- Authors
- VanLeuven, A.J., Park, S., Menke, D.B., Lauderdale, J.D.
- ID
- ZDB-PUB-180626-6
- Date
- 2018
- Source
- Biotechniques 64: 275-278 (Other)
- Registered Authors
- Lauderdale, James D.
- Keywords
- Danio rerio, genotyping, polymerase chain reaction (PCR)
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems/genetics*
- DNA/analysis
- DNA/genetics
- Genotyping Techniques/methods*
- Mutation/genetics*
- Native Polyacrylamide Gel Electrophoresis/methods*
- Polymerase Chain Reaction/methods*
- Zebrafish/genetics*
- PubMed
- 29939088 Full text @ Biotechniques
Citation
VanLeuven, A.J., Park, S., Menke, D.B., Lauderdale, J.D. (2018) A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafish. Biotechniques. 64:275-278.
Abstract
The introduction of CRISPR-Cas9 technology for targeted mutagenesis has revolutionized reverse genetics and made genome editing a realistic option in many model organisms. One of the difficulties with this technique is screening for mutations in large numbers of samples. Many screening approaches for identifying CRISPR-Cas9 mutants have been published; however, in practice these methods are time consuming, expensive, or often yield false positives. This report describes a PCR-based screening approach using non-denaturing PAGE. This approach does not depend on the formation of heteroduplexes and reliably detects changes as small as 1 base-pair (bp) in nucleic acid length at the target site. This approach can be used to identify novel mutations and is also useful as a routine genotyping method.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping