Team:AFCM-Egypt/Improve





We have created 14 parts with functional improvements for the purpose of fitting into lentiviral transfection-required vectors, we have selected one of these parts for judging evaluation and collected them into a one-part collection for the specific purpose of lentiviral delivery of mIRNAs. All these parts have been submitted to IGEM registry of parts and provided with essential characterization on their registry pages.




<p>1- We have managed to improve AmpR at  BBa_K2308011 by fusing AmpR to AmpR promoter which was found to be RhoN and constitutively essential for plasmid selection in lentiviral delivery transfer plasmids. This part has been submitted after improvements to this composite Part BBa_K2534007 .

2- We have provided specific characterization for BBa_K2534018 by using eGFP for experimental characterization of lentiviral transfer plasmid transfection efficacy and apoptotic effect on colorectal cancer cell line RKO. This part is also an improved version of EGFP at [http://parts.igem.org/Part:BBa_I714891# BBa_I714891] by composting the kozak sequence into it for purposes of lentiviral delivery.




3- We have created BBa K2534022 tetracycline Resistance gene as an efficient switching control system for lentiviral delivery of mIRNAs into Colorectal cancer cell line RKO. This comes in a trial to to improve tetR registry part BBa_K182005 which lacks a terminator We also worked to improve this part by adding a universal terminator To improve the control of the highly specific interaction of the tet repressor protein (TetR) with (tetO). TetR binding on tetO classically stops expression of genes downstream of tetO by excluding RNA polymerase from binding the promoter and initiating transcription.




34- we also improved this part BBa_K1119006 on BBa K2534023 by Fusing CMV enhancer to CMV promoter in one composite part to enhance its transcription activity.[https://2018.igem.org/Team:AFCM-Egypt# Team:AFCM-Egypt 2018] have characterized this part considering it's construction into the transfer vector of the lentiviral delivery.br />