Rice thousand kernel weight gene TGW6 guided RNA target sequence and application thereof
A TGW6U3-T1-R, target sequence technology, applied in the field of plant biology, can solve the problem of lack of evaluation of the value of related mutants, and achieve the effect of efficient editing
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0041] Example 1 Creation of rice thousand-grain weight gene tgw6 deletion mutant based on CRISPR / Cas9 technology
[0042] (1) Design of guide RNA (guideRNA, gRNA) target sequence
[0043] According to the genome sequence of the rice grain length and thousand-grain weight gene TGW6 (GenBank: AB513135.1), three gRNAs targeting the thousand-grain weight gene TGW6 were designed. The 20nt oligonucleotide gRNA target sequence is designed according to the A / G(N)20NGG sequence, and the designed gRNA target sequence is compared with the rice genome database to exclude non-specific target cutting sites. The nucleotide sequence is shown in Table 1 and figure 1 .
[0044] Table 1 Oligonucleotide sequences of gRNA targets
[0045]
[0046] (2) Construction of three-target CRISPR / Cas9-gRNA vector
[0047] Referring to the method of Ma et al. (MaX, ZhangQ, ZhuQ, LiuW, ChenY, QiuR, WangB, YangZ.2015.ArobustCRISPR / Cas9systemforconvenient, high-efficiencymultiplexgenomeeditinginmonocota...
Embodiment 2
[0059] (1) Agrobacterium-mediated genetic transformation of rice callus
[0060] The CRISPR / Cas9-gRNA vector assembled in Example 1 was transformed into Agrobacterium EHA105 by electric shock. PCR detection (primers: hptF: 5'-TCCGGAGCCTCCGCTCGAAGTAG-3', hptR: 5'-CTGAACTCACCGCGACGTCTGTC-3') positive clones were used to infect rice material H447 (BC of R819 / Yuzhenxiang / / R819 2 f 7 ), the infection method was carried out with reference to the method of Hiei et al. (1994), and transgenic rice plants were obtained.
[0061] (2) Extraction of rice genomic DNA and PCR detection and sequencing analysis of mutants
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com