Highly efficient CRISPR/Cas9 genome editing in Caenorhabditis elegans has been facilitated by using single-stranded oligonucleotide donors as repair templates. However, insertion of larger sequences remains challenging. Here, Matthew Eroglu, Bin Yu and Brent Derry simplified the generation of long ssDNA as donors in CRISPR/Cas9. High ssDNA yields are rapidly generated using a standard PCR followed by an enzymatic digest with lambda exonuclease. The insertion frequency for these long ssDNA donors is remarkably higher than dsDNA. This can be leveraged to simultaneously generate multiple large insertions and successful edits. This approach enables highly efficient insertion of longer sequences using CRISPR/Cas9 in C. elegans.

We investigated the half-lives of miRNAs in adult skeletal muscle with an in vivo metabolic labeling approach and a genetic mouse model. In vivo metabolic labeling and chasing of miRNAs revealed that the half-lives of skeletal-muscle-enriched miRNAs were 11–20 h. Furthermore, the miR-23a levels decreased rapidly in the skeletal muscle of mice with an inducible loss of miR-23 clusters. These findings suggest that the half-lives of miRNAs were relatively short.

The post-translational process O-GlcNAcylation is implicated in a variety of physiological processes and diseases, including hepatocellular carcinoma. Here, we identified HLY838, a novel diketopiperazine-based OGT inhibitor that induces a global decrease in cellular O-GlcNAcylation. HLY838 enhances the anti-HCC activity of CDK9 inhibitors both in vitro and in vivo by downregulating c-Myc and E2F1 expression. Our findings suggest that targeting O-GlcNAcylation with HLY838 may be an effective strategy to improve cancer therapy.

The liver is one of the few organs that retain the capability to regenerate in adult mammals. Hepatocytes play a crucial role in liver regeneration through reprogramming and expansion after injury. Here, we review the molecular mechanisms governing the remarkable plasticity of hepatocytes in various liver disorders, with a focus on injury-induced reprogramming, and highlight its potential therapeutic applications in chronic liver diseases and liver cancer.

Patulin synthase (PatE) from Penicillium expansum is a flavin-dependent enzyme that catalyses the last step in the biosynthesis of the mycotoxin patulin. The patE gene was expressed in Aspergillus niger allowing purification and characterization of PatE. This confirmed that PatE is active not only on the proposed patulin precursor ascladiol but also on several aromatic alcohols including 5-hydroxymethylfurfural. By elucidating its crystal structure, details on its catalytic mechanism were revealed.

Cellular senescence, a critical hallmark of ageing, has been shown to drive many age-associated chronic diseases. Senescent cells (SnCs) up-regulate pathways associated with evasion of apoptosis, survival, senescence-associated secretory phenotype (SASP) development and others that are targeted using senotherapeutics. Senotherapeutics selectively target SnCs for their clearance (senolytics) or suppression of their SASP (senomorphics). Many studies have demonstrated the benefit of senotherapeutic treatment for the extension of health and lifespan.

The inner blood–retina barrier (iBRB) refers to the properties of endothelial cells associated with the blood vessels of the inner retina. These cells strictly regulate entry and exit from the retina and perturbations in their function have detrimental effects on vision and lead to conditions such as diabetic retinopathy (DR) and age-related macular degeneration (AMD). This review provides a succinct overview of BRB breakdown-associated retinal conditions and the putative underlying mechanisms of disease.

Alzheimer’s disease (AD) is a neurodegenerative disease with multifactorial etiology, intersecting risk factors, and a lack of disease-modifying therapeutics. In this review, converging clinical evidence defining lipid dyshomeostasis in early stages of AD is summarized followed by discussions on mechanisms by which lipid metabolism contributes to pathogenesis and modifies disease risk. Furthermore, existing and potential lipid-targeting therapeutic strategies are reviewed.

Mitochondria play a central role in the development of cellular senescence. Senescence is characterized by several mitochondrial functional changes such as a decrease in OXPHOS, reduced levels of NAD⁺ and ATP, and accumulation of TCA cycle metabolites, DAMPs, and ROS. Here, we provide an overview of the recent findings demonstrating how these mitochondrial changes can contribute to the senescence-associated growth arrest and the SASP.

The accumulation of un- or misfolded proteins can lead to the formation of amorphous or ordered aggregates. Protein aggregation is often associated with human diseases and unravelling its underlying mechanism is critical for the development of diagnostic methods or treatments. However, investigating protein aggregation is challenging due to its complex and dynamic nature. Here, we summarized some popular methods to study the various aspects and steps involved in protein aggregation.

Nrf2 activates different cellular mechanisms in response to ischemia-reperfusion (IR) injury in liver. IR is a major cause of graft loss and dysfunction in clinical transplantation and organ resection. During the IR process, ROS generation leads oxidative stress, inflammation and mitochondrial dysfunction. After Nrf2 activation, the downstream antioxidant upregulation can act as a primary cellular defense and help to promote hepatic recovery during IR.

Ferroptosis is regulated by two transcription factors, NRF2 and BACH1, as its suppressor and promoter respectively. Here, we reviewed how these factors regulate execution of ferroptosis by changing ferroptosis-related metabolites (labile iron, glutathione and ubiquinol), lipid metabolism and cell differentiation. Ferroptosis is elaborately regulated by the competition of NRF2 and BACH1. Death signal is secreted from ferroptotic cells to surrounding cells, and its nature is also discussed.

Cellular senescence functions as a tumour suppressive mechanism, inhibiting the proliferation of cancer-prone cells. In precancerous cells, major autocrine/paracrine tumour-suppressive effects promote senescence surveillance by immune cells. However, in advanced cancer, the altered secretory profile of senescent cells, termed senescence-associated secretory phenotype (SASP), enables the formation of tumour-promoting microenvironments. While SASP and its effects on tumorigenesis are context dependent, deciphering the mechanisms underlying such diversity will allow targeting SASP effectively for cancer therapy.

Following the ground-breaking discovery of imatinib for the treatment of chronic myeloid leukaemia, there has been increasing interest in harnessing kinases as drug targets for cancer. In this article, we provide an update on the current progress with small-molecule kinase inhibitors that have been approved for clinical use. We then discuss the application of mass spectrometry-based proteomics strategies as well as the challenges and outlook for kinase inhibitor research.

MYC and BRD4 play central roles as chromatin and transcriptional regulators in controlling growth, development, stress responses, metabolism and genome stability. This review discusses MYC and BRD4 regulation, their overlapping roles in various cellular processes and how their functional or regulatory impairment leads to disease. Understanding this complex interplay between the two proteins can inform treatment modalities that will effectively counter their pathological effects while preserving normal cellular function.