| 1. |
- Singh, K., et al.
(författare)
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Deep eutectic solvents based biorefining of Value-added chemicals from the diatom Thalassiosira andamanica at room temperature
- 2022
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Ingår i: Separation and Purification Technology. - : Elsevier BV. - 1383-5866. ; 298
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Tidskriftsartikel (refereegranskat)abstract
- The extraction and biorefining of value-added chemicals from bioresources using green solvents are among the key agenda of the circular bioeconomy. Herein we have developed a deep eutectic solvent (DESs) based biorefining approach for clean separation and purification of value-added Fucoxanthin, Chlorophyll, and Biosilica from a diatom microalgae, Thalassiosira andamanica. Several hydrophilic/hydrophobic (DESs) based on quaternary ammonium salt as hydrogen bond acceptor and organic acids and alcohols as hydrogen bond donors were synthesized and tested for microalgae biomass dissolution and biorefining via the phase partitioning method. In an optimized process, similar to 30 wt% of fresh weight diatom could be dissolved in hydrophilic DESs at room temperature, followed by a record extraction of 19.93 mg/g of Fucoxanthin via phase partitioning, with a 53 % increase in yield and an 81 % increase in selectivity over conventional solvents. H-1 NMR, FTIR, LCMS, UV, and HPLC techniques were used to characterize the extracted Fucoxanthin. Subsequently, a hydrophobic DES was added to the remaining hydrophilic DESs and residual biomass and Chlorophyll were extracted into the upper hydrophobic DES layer and Biosilica (80 mg/g) was recovered by simple centrifugation. Extracted Biosilica showed an excellent adsorption capacity of 224.71 mg/g for the removal of methylene blue (MB) dye from water (94.3% up to 88.8 % after 4th consecutive steps). Whereas Chlorophyll extracted in the hydrophobic DES phase exhibited excellent photostability (6 folds greater than conventional solvent) indicating the efficacy of DESs as photoprotector for light-sensitive pigments. This is the first report wherein besides an extractant DES also acted as a good photostabilizer. Additionally, we have replaced the acid-based extraction method for Biosilica with non-toxic DESs.
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| 2. |
- Mandal, Pankaj K, et al.
(författare)
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Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9.
- 2014
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Ingår i: Cell Stem Cell. - : Elsevier BV. - 1934-5909. ; 15:5, s. 643-652
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Tidskriftsartikel (refereegranskat)abstract
- Genome editing via CRISPR/Cas9 has rapidly become the tool of choice by virtue of its efficacy and ease of use. However, CRISPR/Cas9-mediated genome editing in clinically relevant human somatic cells remains untested. Here, we report CRISPR/Cas9 targeting of two clinically relevant genes, B2M and CCR5, in primary human CD4(+) T cells and CD34(+) hematopoietic stem and progenitor cells (HSPCs). Use of single RNA guides led to highly efficient mutagenesis in HSPCs but not in T cells. A dual guide approach improved gene deletion efficacy in both cell types. HSPCs that had undergone genome editing with CRISPR/Cas9 retained multilineage potential. We examined predicted on- and off-target mutations via target capture sequencing in HSPCs and observed low levels of off-target mutagenesis at only one site. These results demonstrate that CRISPR/Cas9 can efficiently ablate genes in HSPCs with minimal off-target mutagenesis, which could have broad applicability for hematopoietic cell-based therapy.
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