Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis
Research output: Contribution to journal › Journal article › Research › peer-review
CRISPR/Cas9 system directed by a gene-specific single guide RNA (sgRNA) is an effective tool for genome editing such as deletions of few bases in coding genes. However, targeted deletion of larger regions generate loss-of-function alleles that offer a straightforward starting point for functional dissections of genomic loci. We present an easy-to-use strategy including a fast cloning dual-sgRNA vector linked to efficient isolation of heritable Cas9-free genomic deletions to rapidly and cost-effectively generate a targeted heritable genome deletion. This step-by-step protocol includes gRNA design, cloning strategy and mutation detection for Arabidopsis and may be adapted for other plant species.
Original language | English |
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Article number | 3796 |
Journal | Bio-protocol |
Volume | 10 |
Issue number | 20 |
Number of pages | 17 |
ISSN | 2331-8325 |
DOIs | |
Publication status | Published - 2020 |
- CRISPR/Cas9, Cas9-free, dual-sgRNA, gBlock, Genomic deletion, FUNCTIONAL GENOMICS, GENE DELETION, CRISPR/CAS9, MUTATIONS, SYSTEM, DNA
Research areas
Links
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7842341/pdf/BioProtoc-10-20-3796.pdf
Final published version
ID: 251575278