Team:IISER Bhopal/Engineering

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Engineering Success

Introduction

Our kill switch characterization demonstrates our engineering success and the results for the same are presented in the results. You can also view the description, mechanism, modelling and results of the kill switch in the registry part page - Part:BBa_K3848004.

Unfortunately due to the COVID-19 pandemic, we had delayed lab access and inconsistent deliveries by the vendors causing our core project lab work to begin in September, hence we were not able to execute all of our modules. But we were successfully able to demonstrate the engineering success of our kill switch.

All the protocols mentioned here are described in the protocols page, here we present the workflow of the project. In the workflow, protocols described in the protocols page are highlighted.

MODULE 1

Initial Characterization:-

Bacterial Growth Kinetics ‎‎‎‎‎‎‎ ‎‎‎

Transformation of bacteria with pUC 19, pMB1

Extraction of Plasmid and Checking for plasmids pMB1

PCR To amplify the ordered gene constructs

MODULE 2

Cloning & Expression

Restriction Digestion ‎‎‎‎‎‎‎ ‎‎‎

Sequential ligation

Transformation, Selection

AND Gate Characterization

Kill Switch Characterization

TRAIL-Smac FP expression

MODULE 3

Cell Line Studies

Cell line Culture ‎‎‎‎‎‎‎ ‎‎‎

TRAIL Binding Assay

Cleavage of Fusion Protein

Internalization of Smac

Apoptosis Detection Assays

Mechanism of Apoptosis

MODULE 4

Future Aspects

B. longum studies ‎‎‎‎‎‎‎ ‎‎‎

Animal Model Studies

Module 5

Proposed implementation

Our product Trasmax to be administered as IV injection is a pharmaceutical product and our end-users are cancer patients. Our target group mainly includes early stage and pre- metastasis stage cancer patients.

This novel treatment therapy can be used to treat cancer patients at a much cheaper cost with minimal side-effects. Though we do not have accurate wet-lab data to support this claim, as per our understanding from previous literature reviews, B.longum elicited minimal immune response when tested on Guinea pigs.

We have completed a thorough analysis of the market (how we intend to reach and diversify our market), competitors, customer needs, requirements (the assistance we can give to the diabetes patients), patient demographics etc. These are just some of the various aspects we have clearly identified and explained in the entrepreneurship section of Implementation. Our safety policies are extremely stringent which will not only ensure the safety of the patient but also that of the environment. We have rigorously tested for biosafety issues and tried to eliminate them in the design stages.