Tomato u6 promoter
WebSep 5, 2024 · The PCR fragment was amplified using the plasmid pSG-SlU6 (Biogle GeneTech, Jiangsu, China) as a template to generate the PCR product containing the Bsa I site, two targets, gRNA scaffold, and the tomato U6 promoter (Fig. S4B, see online supplementary material). WebTomato (Solanum lycopersicum) ... under control of the Arabidopsis U6 polymerase promoter are assembled into a T-DNA vector. Generally, we design CRISPR/Cas9 …
Tomato u6 promoter
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WebFeb 9, 2024 · CRISPR-Cas9 vector construction. In the CRISPR-Cas9 system we used, the sgRNA is transcribed by the Arabidopsis U6 promoter (P U6) and the Cas9 gene is expressed ectopically by the 35S promoter (Fig. 2).An Arabidopsis codon optimized Cas9 protein is used in this system [].The backbone of the destination vector is the pMDC32 … WebMar 8, 2024 · To construct vector pPE2-35SRat, NLS, U6 promoter, double 35S promoter, and nCas9(H840A) codon-optimized for Arabidopsis by Tsingke, Shanghai, China, the engineered M-MLV reverse transcriptase which was amplified from Addgene# 132775, Cambridge MA, USA, EGFP which was from Prof. Gong, and T2A linker were cloned into …
WebJul 21, 2015 · Similar results were reported in tomato ... U6-26p and U6-29p, two Arabidopsis U6 gene promoter; U6-26t, U6-26 terminator with downstream sequence; Hyg, hygromycin-resistance gene. For the sgRNAs, the yellow part represents 20-bp target and the green part represents 76-bp sgRNA scaffold. ... Web【Objective】U6 promoter is a vital element for the transcription of sg RNA in the CRISPR/Cas9 system. It is necessary to clone some endogenous U6 promoters with high transcription activity and construct CRISPR/Cas9 vector, which could provide a strong technical basis for functional genomics and molecular breeding in tomato. 【Method】 …
WebThe corresponding crRNA is expressed using a PCR fragment containing a U6 promoter fused to the crRNA sequence. Transfected cells were analyzed using either Surveyor nuclease assay or targeted deep sequencing. FIG. 101B (top) depicts the sequence of DNMT1-targeting crRNA 3, and sequencing reads (bottom) show representative indels. IG. WebApr 10, 2024 · The MOSAICS AAV vector carries a U6 promoter driven sgRNA element and a CAG promoter driven Sleeping Beauty 100 transposase (SB100)-P2A-Cre fusion protein (Figure 2A). The entire AAV payload is flanked by transposon inverted repeat (IR) sequences that enable genome integration of the payload mediated by SB100 ( Figure 2 …
WebJan 15, 2024 · This vector contains an Arabidopsis thaliana U6 promoter driving the expression of the gRNA scaffold and a BsaI restriction site downstream to enable cloning of the gRNAs. Three guide RNAs were designed as complementary pairs of DNA oligos, annealed to form double-strand fragments, and then individually cloned at the Bsa I …
Web1. A plant or part thereof comprising at least one mutation in an endogenous Short Internodes (SHI) transcription factor gene that encodes a SHI transcription factor comprising a zinc-finger DNA binding domain (ZnF domain), wherein the mutation disrupts the binding of the SHI family transcription factor to DNA, optionally wherein the SHI transcription … i\u0027m and out torranceWebJan 16, 2024 · Result 4 kinds and 8 different lengths of tomato U6 promoters were obtained by two rounds of PCR. Their length was 452, 202, 448, 206, 433, 190, 448 and 218 bp, … net marginal product of capitalWebJun 9, 2024 · The U6 promoter with +1A/G efficiently produces small RNA by precise +1 transcription. The human U6 promoter controls the transcription of the U6 small nuclear RNA (snRNA), which is an essential component of the spliceosome. Citation 33 The transcription start site (+1G) of the human U6 snRNA is located 24-nt downstream of the … net margin industry benchmark dataWebU6 prom Rv: GGTGTTTCGTCCTTTCCAC Use the purified U6 promoter as a template and set up the following reaction (set up the reaction 5x or more to get enough product): 5x … net margins by industryWebAug 14, 2015 · zt176 PiggyBac Dual promoter (PB513B-1) 载体 SBI PiggyBac 转座子载体系统. zt177 Super PiggyBac Transposase (PB200PA-1) 载体 SBI PiggyBac 转座子载体系统. zt178 信号通路报告载体系统： 信号通路报告载体系统： zt179 pGAS-TA-Luc 载体 Clontech 信号通路报告载体系统： i\\u0027m an educated fool with money on my mindWebRecently, the CRISPR/Cas9‐mediated targeted mutagenesis of GmSPL9 , which were expressed using Arabidopsis U3 and U6 promoters, ... In tomato, a novel mechanism of interspecific incompatibility was recovered by knockout of … net margin analysisWebConstructs alternating sgRNA and tRNA sequences under the control of a single U3 or U6 promoter permit reduction of the size of the construct and limit the risk of silencing due to direct repetitions of promoter ... SgRNAs were specific for different tomato yellow leaf curl virus (TYLCV) coding and non-coding sequences, targeting the viral ... net margin for a retailer