Search

Collaborative governance is increasingly being used as a solution to address climate change and deforestation in the tropics, but its stakeholders face numerous challenges in making it work. This study aims to understand stakeholders’ perspectives on the challenges of collaborative governance, focusing on Sabah’s jurisdictional approach. We applied the Q-methodology to derive the perspectives of the stakeholders involved. The results revealed three significant perspectives. The first perspective, “participant factors,” highlighted that the representation of the ‘right’ stakeholders and the mandate to make decisions are inadequate. The second perspective, “non-progress in activities,” suggests that the lack of accountability hampers the progress of the initiative. The third perspective, “shared understanding,” reflects the stakeholders’ inability to agree on a common goal. The consensus regarding the collaboration challenge is that the jurisdictional approach initiative is new, and nobody knows how to implement it, emphasizing the need for higher-level government commitment. This study reveals the challenges of collaborative governance in a jurisdictional approach by providing empirical evidence of the diverse perspectives of stakeholders.

In multicellular organisms, regulation of telomere length in pluripotent stem cells is critical to ensure organism development and survival. Telomeres consist of repetitive DNA that are progressively lost with each cellular division. When telomeres become critically short, they activate a DNA damage response that results in cell cycle arrest. To counteract telomere attrition, pluripotent stem cells are equipped with telomere elongation mechanisms that ensure prolonged proliferation capacity and self-renewal capacity. Excessive telomere elongation can also be deleterious and is counteracted by a rapid telomere deletion mechanism termed telomere trimming. While the consequences of critically short telomeres are well established, we are only beginning to understand the mechanisms that counteract excessive telomere elongation. The balance between telomere elongation and shortening determine the telomere length set point in pluripotent stem cells and ensures sustained proliferative potential without causing chromosome instability.

Seamless cloning methods, such as co-transformation cloning, sequence- and ligation-independent cloning (SLIC) or the Gibson assembly, are essential tools for the precise construction of plasmids. The efficiency of co-transformation cloning is however low and the Gibson assembly reagents are expensive. With the aim to improve the robustness of seamless cloning experiments while keeping costs low, we examined the importance of complementary single-stranded DNA ends for co-transformation cloning and the influence of single-stranded gaps in circular plasmids on SLIC cloning efficiency. Most importantly, our data show that single-stranded gaps in double-stranded plasmids, which occur in typical SLIC protocols, can drastically decrease the efficiency at which the DNA transforms competent E. coli bacteria. Accordingly, filling-in of single-stranded gaps using DNA polymerase resulted in increased transformation efficiency. Ligation of the remaining nicks did not lead to a further increase in transformation efficiency. These findings demonstrate that highly efficient insert-plasmid assembly can be achieved by using only T5 exonuclease and Phusion DNA polymerase, without Taq DNA ligase from the original Gibson protocol, which significantly reduces the cost of the reactions. We successfully used this modified Gibson assembly protocol with two short insert-plasmid overlap regions, each counting only 15 nucleotides.