Biology subjects

Manvati, S.

Publications and source records attributed to Manvati, S..

4 recordsLinked to original sources

bioRxiv2023

Harnessing Escherichia coli Dark Genome to Produce Anti-Alzheimer Peptides

Alzheimers disease (AD) is characterized by progressive neurodegeneration. The critical molecular trigger is believed to be the accumulation of A{beta} neurotoxic oligomers. Given the proteolytic processing of Amyloid Precursor Protein (APP) by {beta}-secretase (beta-site APP cleaving enzyme 1, BACE1) as the key step in the building up of A{beta} oligomers, BACE inhibitors come with therapeutic prospects of preventing or delaying the onset of Alzheimers. To find inhibitory peptides against BACE1, a library of dark peptides was constructed from 4400 intergenic DNA sequences of Escherichia coli. The sequence level analysis was followed by protein structure predictions, molecular docking, and simulation. Based on bioinformatics analysis, 5 potential peptides were screened for experimental validation. Out of these two peptides were identified as lead molecules based on BACE1 inhibitory activity, followed by FRET inhibitory assay, western blot, and RT-PCR. An 86.7 % drop in BACE1 level was observed in the presence of the ECOI2 peptide. Though encouraging results were obtained from in-silico and in-vitro studies, more work is required to study the efficacy of these peptides in suitable animal models.

synthetic biology↗

bioRxiv2023

ClearX9-Stem™: an efficient, affordable & sustainable stem cell culture medium for biotech and cell-cultivated meat industries

Stem cells are extraordinary cells with a unique ability of self-renewal and differentiation into various cell types such as muscle, nerve, bone, and blood cells. Historically, they have found significant applications in the biotech and pharma sectors. To grow and maintain stem cells artificially, researchers use basal media formulations supplemented with nutrients and growth factors, with Fetal bovine serum (FBS) as the key component of the culture medium. However, to maintain this supply every year, millions of pregnant cows are slaughtered for preparing FBS. The process of harvesting FBS also raises concerns about contamination with pathogens, animal proteins that may interfere with cellular behavior and ethical considerations regarding animal welfare. To overcome these limitations, here we report ClearX9-Stem - an affordable, sustainable, effective, and ethical replacement for an FBS-enriched stem cell culture medium. A specialized ClearX9-Stem cell culture medium formulation was designed to grow chicken embryonic fibroblast (SL-29) in the absence of FBS. Based on the results obtained, ClearX9-Stem is undergoing further refinement to meet the growing academic and industrial demand for serum-free culture media formulations. In the future, there is a need to customize and optimize ClearX9-Stem for the scalable growth of cells in bioreactors. HIGHLIGHTSO_LIClearX9-Stem provides good nutritional support for the growth of chicken embryonic fibroblast cells. C_LIO_LIClearX9-Stem cell growth performance is comparable to the serum-enriched culture medium C_LIO_LIClearX9-Stem maintains a healthy morphological profile of cells during division C_LIO_LIClearX9-Stem generates a stress-free environment within cells C_LIO_LIClearX9-Stem does not require animal slaughter and reduces the environmental footprint C_LIO_LIClearX9-Stem has applications in the biotechnology, pharma, and cell-cultivated meat industries C_LI

synthetic biology↗

bioRxiv2023

ClearX9™: an efficient alternative to fetal bovine serum for growing animal cells in vitro

Fetal Bovine Serum (FBS) is a nutrient-rich fluid that contains nutritional and macromolecular factors essential for cell growth. Every year millions of pregnant cows are slaughtered in search of FBS leading to huge environmental consequences. Here we report ClearX9 - an affordable, sustainable, ethical, and effective replacement for FBS. ClearX9 cell culture medium was used to grow HeLa (cervical cancer cells), HEK293T (embryonic kidney transformed cells) and Nthy Ori-3-1 (primary thyroid follicular transformed epithelial cells) and showed encouraging growth patterns and good cellular health. Compared with the FBS-enriched cell culture medium, ClearX9 scored positive on all the parameters suggesting ClearX9 as a credible alternative to FBS. In future, more work is required to establish the efficacy of ClearX9 in toxicology testing, bio-manufacturing, regenerative medicine, and vaccine research. HIGHLIGHTSO_LIClearX9 provides good nutritional support for the growth of animal cells C_LIO_LIClearX9 cell growth performance is comparable to the serum-enriched medium C_LIO_LIClearX9 maintains a healthy morphological profile of cells during division C_LIO_LIClearX9 generates a stress-free environment within cells C_LIO_LIClearX9 does not require animal slaughter and reduces carbon footprint C_LIO_LIClearX9 has applications in biotechnology and cell cultivated meat industry C_LI

cell biology↗

bioRxiv2020

Discovery of tREP-18, a novel class of tRNA encoded peptide with potent leishmanicidal activity

In the post genomic era, tRNA-derived fragments have emerged as a new class of non-coding gene regulators, those play crucial roles both at the transcriptional and translational levels, in different cellular biogenesis. However, none of the studies has ever asked whether tRNAs can also be translated into peptides with any biological significance. Thus, we present a novel hypothesis which suggested that; design and synthesis of tRNA-derived peptides from prokaryotic genome can be exploited for developing unique chemotherapeutics against neglected tropical diseases, like Visceral leishmaniasis (VL) and its aggressive form known as post kalazar dermal leishmaniasis (PKDL). To achieve this aim, we have used a novel system biology-based strategy, which involved; i) mining of unique tRNAs from E. coli genome and their translation into peptide in silico, ii) designing of theoretical 3D models to evaluate their stability, iii) prediction of their biological activity by screening against anti-parasitic database to filter the lead peptide. Based on this strategy, a unique tRNA-derived peptide (tREP-18) was selected, chemically synthesized, and used in vitro for elucidating its therapeutic significance against L. donovani, a causative agent of VL and PKDL. Our findings demonstrated that, tREP-18 can impose high level toxicity to L. donovani promastigotes, by disrupting the ultrastructural cellular architect, destabilizing the mitochondrial membrane potential ({Delta}{Psi}m), thus leading to drastic reduction in cell viability and proliferation. It also imparted high level of toxicity to BS12 a clinical isolate of PKDL. Conceivably, we for the first time reports a novel tRNA-derived peptide "tREP18" with excellent anti-leishmanial property, which can be further utilized for developing it as antileishmanial drug. One Sentence SummaryHere we report a novel first-in-class tRNA encoded peptide and its super anti-leishmanial characteristics.

synthetic biology↗