Phee401e vector
WebMar 15, 2024 · Then the fragment was ligated into the binary vector pHEE401E by the restriction-ligation system ( Wang et al., 2015 ). Then, the recombinant plasmid pHEE401E-2sgRNA-SlNAC063 was transformed into wild-type tomato AC using the stable A. tumefaciens -mediated transformation method ( McCormick et al., 1986; Gupta and Van … WebAug 16, 2024 · Building on the pHEE401E plasmid, a Cambia T-DNA adapted for 96 genome editing with CRISPR/Cas9 nucleases (Wang & al., 2015), we created a series of 97 vectors enabling selection and counter-selection of transgenics on the basis of seed 98 fluorescence. Toward this end, we replaced the hygromycin resistance marker of pHEE401E
Phee401e vector
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WebOct 24, 2024 · Two sgRNAs were designed and cloned into pHEE401E as described by Wang et al. (2015b). ... China) for the CRISPR/Cas9 vector; Xiao-Su Gao, Jiqin Li, Yun-Xiao He, and Shui-Ning Yang (Chinese Academy of Sciences Center for Excellence in Molecular Plant Sciences, China) for skillful technical assistance; Hongtao Liu, Lin Xu (Chinese Academy … WebJul 21, 2015 · Arabidopsis mutants produced by constitutive overexpression of the CRISPR/Cas9 genome editing system are usually mosaics in the T1 generation. In this study, we used egg cell-specific promoters to drive the expression of Cas9 and obtained non-mosaic T1 mutants for multiple target genes with high efficiency. Comparisons of 12 …
WebJun 22, 2024 · a A schematic representation of the gene drive that is inserted into the second exon of the CRYPTOCHROME 1 ( CRY1) gene on Chromosome IV using CRISPR/Cas9-mediated homology-directed repair (HDR).... WebSep 24, 2024 · The structure of CSE-653 vector designed to sequentially cut S653 codon twice. pHEE401E backbone was used to ubiquitously express two sgRNAs by U6-26 and U6-29 promoters, and Cas9 was driven by an ...
Webprimer set and ligation strategy, the pHEE401E vector can carry four sgRNAs, and the pBAtC-tRNA vector can carry four to six sgRNAs. 2. Experimental Design Two main vectors were … WebpHEE401E (Plasmid #71287) Print Enlarge View all sequences Purpose Egg cell-specific promoter-controlled expression of 3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for … Plasmid pHEE401 from Dr. Qi-Jun Chen's lab contains the inserts sgRNA scaffold … Plasmid pCBC-DT1T2 from Dr. Qi-Jun Chen's lab contains the insert T1, U626t …
WebDec 19, 2024 · To construct pHEE401E (Wang et al., 2015) vectors targeting four different sites of PAR1 and one site of JAZ1 or GAI, the dsDNA was fused to the BsaI-digested …
WebPrinciple and design of JMJ epigenome editing constructs targeting APX2. Schematic representation of generated constructs for validation of sgRNA design by transient transactivation in N. benthamiana (A) and for generation of stable lines (B) in the pHEE401E vector backbone. Both types of constructs contain an sgRNA cassette, which is … fowler dallas cowboysWebNov 22, 2024 · The pHEE401E_UBQ_Bar vector is a version in which the egg-specific promoter for Cas9 expression in the pHEE401E vector was replaced with the UBQ10 … black storage furnitureWebAug 13, 2024 · EAR (Ethylene-responsive element binding factor-associated Amphiphilic Repression) motif-containing transcription repressors have been shown to regulate plant growth and development, and plant responses to plant hormones and environmental stresses including biotic and abiotic stresses. However, the functions of most EAR-motif … black storage jars tea coffee sugarWebModified from the pHEE401E T-DNA vector described by Wang &al. (2015) Genome Biol 16: 144, doi:10.1186/s13059-015-0715-0: the Hygromycin selection maker of the T-DNA was exchanged with a cassette enabling selection on the basis of CFP-fluorescence in seeds; the egg-cell specific promoter of Cas9 was exchanged with a ubiquitin promoter. black storage cube boxesWebpHEE401E (~100 ng/μl) 2 μl 10× T4 DNA Ligase Buffer (NEB) 1.5 μl 10× BSA 1.5 μl BsaI (NE B) 1 μl T4 DNA Ligase (HC, NEB) 1 μl ddH 2 O 6 μl Total volume 15 μl 5. Transform E.coli … black storage ottoman benchWebNov 22, 2024 · The pHEE401E vector has a type IIS restriction-enzyme-recognition site for ligation [6]. The other is a pBAtC-tRNA vector that uses a polycistronic tRNA-gRNA … black storage ottoman walmartWebI am trying to synthesize CRISPR constructs, using Golden Gate to put my sgRNAs in the pHEE401E vector. Whenever I do colony PCR with my original primers, it appears that the reaction worked as I... fowler dealership