Repositorium
Journal Article / 2014
Shan, Qiwei; Wang, Yanpeng; Li, Jun; Gao, Caixia
Targeted genome editing nucleases, such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), are powerful tools for understanding gene function and for developing valuable new traits in plants. The clustered regularly interspersed short palindromic repeats (CRISPR)/Cas system has recently emerged as an alternative nuclease-based method for efficient and versatile genome engineering. In this system, only the 20-nt targeting sequence within the single-guide RNA (sgRNA) needs to be changed to target different genes. The simplicity of the cloning strategy and the few limitations on potential target sites make the CRISPR/Cas system very appealing. Here we describe a stepwise protocol for the selection of target sites, as well as the design, construction, verification and use of sgRNAs for sequence-specific CRISPR/Cas-mediated mutagenesis and gene targeting in rice and wheat. The CRISPR/Cas system provides a straightforward method for rapid gene targeting within 1-2 weeks in protoplasts, and mutated rice plants can be generated within 13-17 weeks.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
424 |
Gao, Caixia China |
Oryza sativa |
CRISPR/Cas9 DEP1 |
Grain number/ grain yield/ plant hight/ spukelet number SDN1 |
Basic research Basic research |
425 |
Gao, Caixia China |
Oryza sativa |
CRISPR/Cas9 MPK2 |
ATP binding/ MAP kinase activity/ protein tyrosine kinase activity SDN2 |
Basic research Basic research |
426 |
Gao, Caixia China |
Oryza sativa |
CRISPR/Cas9 PDS |
Albino and dwarf phenotype SDN1 |
Basic research Basic research |
427 |
Gao, Caixia China |
Triticum aestivum |
CRISPR/Cas9 LOX2 |
seed storability SDN1 |
Basic research Basic research |