Life Sciences & Biotechnology
Title : | CRISPR/Cas based metabolome editing and genomics assisted glucosinolate pathway elucidation of the miracle tree Moringa oleifera Lam. |
Area of research : | Life Sciences & Biotechnology |
Principal Investigator : | Dr. Rehna Augustine, College Of Horticulture, Uttar Pradesh |
Timeline Start Year : | 2022 |
Timeline End Year : | 2025 |
Contact info : | rehna.augustine@kau.in |
Equipments : | lyophilizer/ freeze dryer |
Details
Executive Summary : | Moringa oleifera is known as miracle tree or wonder tree due to its immense health benefits. Eventhough cultivated mainly as vegetable, different parts of the tree has been used in folk medicine from ancient times. Moringa extract have the healing properties against diabetes, inflammation, bacterial, viral and fungal infections, arthritis, cardiac disorders, cancer etc. Despite its innumerable health benefits, not much molecular work has been carried out in the crop for its further improvement and utilization as a great resource for pharmaceutical and cosmetic industries. Glucosinolates (GSL) are bio-active secondary metabolites present in moringa with very high anticancer activity. For successful metabolic engineering, data on the chemical profile, biosynthesis genes involved in the process, their regulation etc. needs to be investigated. Hence, the proposed research work is an effort towards dissecting the molecular mechanism of moringa GSL biosynthesis and its further manipulation using advanced techniques like transcriptomics, metabolomics and CRISPR/Cas mediated metabolic editing. Objectives 1. GSL profiling at different developmental stages and parts of moringa accessions using HPLC 2. Transcriptome analysis and identification of GSL biosynthesis genes from M. oleifera and validation of the gene expression using real-time RT-PCR 3. Development of cell suspension cultures of M. oleifera and validation of selected GSL biosynthesis genes using CRISPR/Cas system Methodology Different moringa accessions will be collected and the samples will be lyophilized and GSLs will be extracted and analysed using HPLC as described by Augustine et al. (2013). GSL content at different growth stages and organs will be determined. Transcriptome analysis of potential moringa accession will be carried out. Genes related to GSL and sulfur metabolism will be identified. The expression of candidate genes will be validated using qRT-PCR. Moringa cell suspension culture will be developed. Potential genes identified from the study will be targeted for editing using CRISPR/Cas system. CRISPR based activation/ knock-out of the key regulatory genes will be attempted in callus cultures. Molecular and chemotypic analysis will be performed in the edited lines Significance Food security is declining and malnutrition and life style diseases like cancer, gastritis and other infectious diseases are increasing. To diversify the global food supply, overcome malnutrition, and to find new curative medicines to tackle life style diseases, it is necessary to diversify and focus more on crop plants that are underutilized. Identification and further exploitation of GSL genes using the next generation metabolic editing tool, CRISPR,Cas, to enhance the production of bioactive compounds using cell suspension cultures is possible. Large scale production of GSL will be possible on commercial scale which can find use in pharma-nutraceutical industries. |
Total Budget (INR): | 26,62,000 |
Organizations involved