Template Vector (BspQ I) for T7 mRNA Synthesis

Description

The prelinearized template vector contains a T7 promoter, transcription start sequence AGG , 5-untranslated region UTR , 3-UTR, poly A sequence, and BspQ I digestion site, which allows the seamless preparation of an in vitro transcription IVT template. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back The length of additional base pairs scars stemming from the poly A tail directly correlates with the translation efficiency.Three types of DNA templates encoding Firefly Luciferase FLuc mRNA pFLuc-142A: BspQ I Vector with a 142 bp of poly A Cat. # 6146 , pFLuc-102A: BspQ I Vector with a 102 bp of poly A not sold , and pFLuc-105A: Hind III vector with a 105 bp of poly A part of Cat. # 6143 were digested with specific restriction enzymes as indicated. The linearized templates, with differing numbers of additional base pairs downstream of the poly A tail as shown in parenthesis, were then employed in an in vitro transcription process with CleanCap. The resulting in vitro transcribed mRNA was transfected into HEK293T cells to assess the efficiency of mRNA translation, which was quantified by FLuc measurements. The DNA template digested by BspQ I enzyme, lacking any extra sequence at the 3 end of the poly A tail, exhibits the highest capacity for protein translation. Back The length of poly A tail positively affects the translation efficiency.Two types of DNA templates encoding Firefly Luciferase FLuc mRNA, pFLuc-142A: BspQ I Vector with a 142 bp of poly A Cat. # 6146 and pFLuc-102A: BspQ I Vector with a 102 bp of poly A not sold , were digested with BspQ I restriction enzyme. The linearized templates were then employed in an in vitro transcription process with CleanCap. Subsequently, the resulting in vitro transcribed mRNA was transfected into HEK293T cells to assess the efficiency of mRNA translation, quantified by FLuc measurements. The DNA template with a longer poly A tail Cat. # 6146 exhibits the higher capacity for protein translation.