RNA-guided genome engineering: paradigm shift towards transposons.

CRISPR-Cas systems revolutionized the genome engineering field but need to induce double-strand breaks (DSBs) and may be difficult to deliver due to their large protein size. Tn7-like transposons such as CRISPR-associated transposons (CASTs) can be repurposed for RNA-guided DSB-free integration, and obligate mobile element guided activity (OMEGA) proteins of the IS200/IS605 transposon family have been developed as hypercompact RNA-guided genome editing tools. CASTs and OMEGA are exciting, innovative genome engineering tools that can improve the precision and efficiency of editing. This review explores the recent developments and uses of CASTs and OMEGA in genome editing across prokaryotic and eukaryotic cells. The pros and cons of these transposon-based systems are deliberated in comparison to other CRISPR systems.

Enhanced Calvarial Bone Repair Using ASCs Engineered with RNA-Guided Split dCas12a System that Co-Activates Sox 5, Sox6, and Long Non-Coding RNA H19.

Healing of large calvarial bone defects remains challenging. An RNA-guided Split dCas12a system is previously harnessed to activate long non-coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow-derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose-derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However, H19 activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co-activate other chondroinductive transcription factors (Sox5, Sox6, and Sox9) to synergistically potentiate differentiation. It is found that co-activation of H19/Sox5/Sox6 in ASCs elicited more potent chondrogenic differentiation than activation of Sox5/Sox6/Sox9 or H19 alone. Co-activating H19/Sox5/Sox6 in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration.

Preparation of polysubstituted imidazoles using AC-SO3H/[Urea]7[ZnCl2]2 as an efficient catalyst system: a novel method, and α-glucosidase inhibitor activity.

Deep eutectic solvents (DESs) act as both an organic solvent and a useful catalyst for organic synthesis reactions, especially the synthesis of heterocyclic compounds containing the element nitrogen. DESs exhibit many important properties namely large liquid fields, biodegradability, outstanding thermal stability, and moderate vapor pressure. Amorphous carbon-bearing sulfonic acid groups (AC-SO3H) are one of the new-generation solid acids showing strong acid activity. Based on the simultaneous presence of acidic functional groups such as carboxylic acid, phenolic, and sulfonic acid groups, they exhibit many important activities namely strong Brønsted acid, high surface area, high stability, reusability, and recyclability. In this study, AC-SO3H was made from rice husk via the carbonization and sulfonation processes, and the surface properties and structure were examined using contemporary methods such as FT-IR, P-XRD, TGA, SEM, and EDS. And, [Urea]7[ZnCl2]2 was synthesized from urea and ZnCl2 with a mole ratio of 7 : 2; the structure is defined using FT-IR and TGA. By combining AC-SO3H and [Urea]7[ZnCl2]2 we aim to form an effective catalyst/solvent system for the preparation of polysubstituted imidazole derivatives through the multi-component cyclization reaction from nitrobenzenes, benzil, aldehydes, and ammonium acetate. The major products are obtained with high isolation yields above 60%. To assess the catalyst system's activity, the recovery and reusability of the AC-SO3H/[Urea]7[ZnCl2]2 system were examined with hardly any performance modification. In an effort to create potential enzyme α-glucosidase inhibitors, several novel polysubstituted imidazoles were created. Five of these compounds showed good enzyme α-glucosidase inhibitor activity. The most effective substances were IMI-13, IMI-15, and IMI-20, with IC50 values that were greater than the acarbose at 16.5, 15.8, and 11.6 µM, respectively - the acarbose (IC50, 214.5 µM) as the positive control.

Synthesis of polyhydroquinolines and propargylamines through one-pot multicomponent reactions using an acidic ionic liquid immobilized onto magnetic Fe3O4 as an efficient heterogeneous catalyst under solvent-free sonication.

A nano-sized Fe3O4-supported Lewis acid ionic liquid catalyst for the synthesis of polyhydroquinolines and propargylamines under ultrasound irradiation has been developed. LAIL@MNP was synthesized from imidazolium chlorozincate(ii) ionic liquid grafted onto the surface of Fe3O4 nanoparticles and evaluated by FT-IR, TGA, SEM, Raman, TEM, ICP-OES, and EDS. The multicomponent synthesis of polyhydroquinolines and propargylamines proceeded smoothly to afford the desired products in high yields. LAIL@MNP can be separated easily from the reaction mixture and reused for several runs without a significant degradation in catalytic activity.