CRISPR-Cas9 knockin of the HBB gene paves the way for a general therapeutic solution for treating hemoglobinopathies.

Ischemia reperfusion (I/R) injury remains a frequent adverse event that accompanies heart trans-plantation. Oxidative stress and aberrant production of free radicals were regarded as the culprit of cell death and tissue damage in post-transplant IR injury. Mst1 has been identified as a mediator of oxidative stress and Nrf2 regulates anti-oxidative enzymes, however, the interaction between Mst1 and Nrf2 anti -oxidative stress pathway remains to be clarified in the event of cardiac IR injury. Herein, the model of ischemia-reperfusion injury in heterotopic heart transplantation mice was firstly established.. We observed that cardiac IR induced upregulation of Mst1 and activation of Nrf2/HO-1pathway in mice receiving heterotopic heart transplantation. Further Cobalt dichloride-induced oxidative stress model of RAW264.7 macrophage cells were then established to mimic cardiac I/R injury, results showed that exposure to CoCl2 induced the upregulation of Mst1 and activation of Keap1/Nrf2 pathway, and genetic ablation of Mst-1 and inhibition of Keap1/Nrf2 pathway aggravated oxidative damage in those cells. Additional in vivo study showed that transfection of Mst1 shRNA spurred ROS generation and worsened cardiac damage in IR mice. Meanwhile, Mst1-KD mice receiving heart transplantation showed markedly downregulation of Nrf2, HO-1 yet upregulation of Keap1, indicating diminished protective effect against tissue damage caused by IR probably owing to the frustration of Keap1/Nrf2 pathway. Taken together, our findings demonstrated the protective effect of Mst1 from cardiac IR injury via triggering Keap1/Nrf2 axis and suppressing ROS generation, which shed light on the promising role of Mst1 in transitional man-agement of IR injury resulted from cardiac transplantation.(c) 2023 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Puberty is the process of physical maturation of an animal with the capability of sexual reproduction. In domestic animals, age at attainment of puberty is an event that contributes significantly to lifetime reproductive efficiency. Anhui white goat is a native Chinese breed, with the characters of early time puberty onset and high fecundity. However, the molecular mechanism of puberty onset has not been well understood. It has been demonstrated leptin act as a critical metabolic cue linking adipose and the onset of puberty. In the present study, we first assessed the morphological changes in adipose tissue at pre-puberty and puberty onset stages of goats, and then determined the role of leptin to activate signaling pathways which modulate the expression of hypothalamic genes involved in reproduction and metabolism. We found that leptin significantly upregulated the LEPR in the adipose tissue of goat at the puberty onset stage. Moreover, we observed that LEPR, JAK2, STAT3 (P<0.05), KISS1 (P<0.05) were also upregulated in the hypothalamic tissue, except NPY, which was downregulated. Taken together, it is, therefore, inferred that probably leptin secreted in the adipose tissue via its receptor leptin receptor through the JAK2/STA3 signal pathways, promote the upregulation of KISS1, which activates TRPC5, and, inhibit the release of NPY, thereby promoting the release of GnRH, thus hastening the onset of puberty in goat. During this process, leptin, LEPR and its signal pathways may act as a link between the adipose and puberty onset in goat. However, further more studies need to be carried out to verify this point.

Here, we report the scalable fabrication of 2i-functionalized micro-pyramid-array (μPyA/+2i) inserts for use in commercial multi-well plates, as the alternative cultivation platform for maintaining long-term self-renewal and pluripotency of multiple mESCs and mouse induced pluripotent stem cells. Relevant evidence including cell morphology characterization increased alkaline phosphatase activity, high expression of mESC self-renewal markers, decreased levels of differentiation-associated markers, and high proportion of self-renewal marker cells are provided. Further studies demonstrated that μPyA/+2i could cause a higher cell density in mESC colony, and induce gene expression changes. Subsequent studies showed that μPyA/+2i can influence the cytoskeleton and promote cell adhesion through Cldn-7 upregulation. In summary, these μPyA/+2i inserts offer flexible and gelatin-free micro-envriomnets to maintain long-term self-renewal and pluripotency of mESCs. Enabled by the microstructured inserst, the facile stem cell manipulation and transfer among culture dishes will broaden stem cells both in routine and translational applications.

Colon cancer ranks among the most prevalent malignant tumors worldwide, with its incidence rising progressively year by year. The identification of novel research tool for therapeutic targets has thus become an urgent priority. Here, we constructed a CS•CBE integrative delivery and editing system via chitosan and cytosine base editor (CBE), which could successfully target and inhibit the phosphorylation of β-catenin. In vitro studies demonstrated that this system achieved a high delivery efficiency of 86.0% and an editing efficiency of 78% in HCT116 cells, and Transwell and wound-healing assays verified that the CBE-β-catenin-HCT116 cells exhibited an increased proliferation rate. Furthermore, a xenograft tumor model was established in nude mice, and we observed that the tumor volume and weight in the experimental group were both significantly increased compared with those in the control group. Subsequently, the candidate genes identified from transcriptomic results, such as AXIN2 and GPR15, which are downstream of β-catenin pathways, validated the feasibility of the CS•CBE system as a research tool. Collectively, our CS•CBE system serves as a valuable research tool for targeted protein modulation, downstream target identification, and probe specific post-translational modifications analysis in colon cancer research.

The genetic basis of high‐altitude hypoxia adaptation remains largely elusive, particularly the role of HBB. Here, we developed an integrative mesh‐nanorod (MN) cell sorting, chitosan nanoparticle (CS) delivery, and in vivo intraosseous (IO) injection (MN•CS•IO) system to functionally investigate HBB in cross‐species high‐altitude hypoxia adaptation. Using this MN•CS•IO platform, we transplanted high/low‐altitude species HBB‐modified lineage‐negative (Lin ⁻ ) cells into murine models, achieving 95.59% transfection efficiency and 8.15% HBB relative expression, and subjected them to simulated and natural Qinghai‐Tibet hypoxia conditions. High‐altitude HBB recipients exhibited downregulated Rragd expression, activated oxygen transport and hematopoietic lineage pathways, improved behavioral performance, and neuroprotection against hypoxia‐induced damage, compared with low‐altitude animals. MN•CS•IO system data demonstrate that HBB determines high‐altitude hypoxia adaptation. For validation, we generated bone marrow chimeras using CD45.1 hematopoietic cells with Pika‐HBB integrated at the Rosa26 locus. The reconstituted mice exhibited consistent hypoxic behavioral and phenotypic alterations with those observed via the MN•CS•IO system, validating the reliability and stability of HBB‐mediated high‐altitude hypoxia adaptation. Together, our findings demonstrate that HBB is a key genetic determinant of high‐altitude hypoxia adaptation, and also introduce a versatile gene‐function validation tool for evolutionary and biomedical research.