Project description:Repeated parallel losses of inflexed stamens in Moraceae: phylogenomics and generic revision of the tribe Moreae and the reinstatement of the tribe Olmedieae (Moraceae)
| PRJEB37667 | ENA
Project description:Resolving generic limits in Cyperaceae tribe Abildgaardieae using targeted sequencing
Project description:The specific genes that distinguish normal fracture healing from abnormal healing or nonunion in humans are unknown. This study was an exploratory investigation of peripheral blood from 2 chronic nonunion patients collected perioperatively (pre/post revision surgery) and at 3 months post revision follow up for comparison to Acutely injured subjects and Healthy volunteer cohorts analyzed separately. We used microarrays to do a global comparison between 2 chronic nonunion patients collected perioperatively (pre/post revision surgery) and at 3 months post revision follow up.
Project description:Surgical revision of infection can involve multiple procedures. Each revision activates a tissue injury response and disrupts the established bacterial biofilm. However, it is not well understood how the bacteria, immune system, and overall tissue environment coordinate their response to revision. Our results show that immunological niches already compromised by infection – such as the bone marrow, lymph nodes, and circulating blood – further upregulate pro-inflammatory programs in response to revision surgery. This enhanced inflammation at the incision site has no effect on reducing bacteria numbers. Instead, it leads to increased expression of virulence factors, enhanced tissue remodeling, and damage including bone osteolysis and muscle fibrosis. Whereas muscle fibrosis appears to resolve 14 days post-revision, osteolysis continues to progress. These findings show that revision surgery negatively impacts all tissues within the surgical site. It is therefore essential to question the risks and benefits of each revision on a case-by-case basis. Further, understanding the timing and tissue changes associated with revision of infected orthopedic sites will help inform the design of additional interventions to minimize tissue damage and maximize bactericidal effects.
Project description:The specific genes that distinguish normal fracture healing from abnormal healing or nonunion in humans are unknown. This study was an exploratory investigation of peripheral blood from 2 chronic nonunion patients collected perioperatively (pre/post revision surgery) and at 3 months post revision follow up for comparison to Acutely injured subjects and Healthy volunteer cohorts analyzed separately. We used microarrays to do a global comparison between 2 chronic nonunion patients collected perioperatively (pre/post revision surgery) and at 3 months post revision follow up.
Project description:Processing bodies (PBs) are dynamic, membraneless organelles consisting of RNAs and proteins. While PB proteins have been extensively characterized, the methods for systematically profiling PB-associated RNAs are limited. To address this, we developed PB-TRIBE-STAMP, a new tool based on two orthogonal RNA editing enzymes. Simultaneously applying APOBEC1-DDX6 and LSM14A-ADAR2dd, PB-TRIBE-STAMP identified 1,639 and 2,577 PB-associated mRNAs in HCT116 and HEK293T cells, respectively. Further biochemical isolation of PBs followed by RNA-seq validated that edited transcripts of these mRNAs were indeed enriched in PBs. Integration of PB-TRIBE-STAMP with long-read sequencing revealed that the PB-associated transcripts possessed shorter poly(A)-tails and mRNA isoforms with longer 3’ UTRs were more likely to be associated with PBs than those with shorter ones. Moreover, we established a TRIBE-ID-based tool to characterize the mRNA-PB association at high temporal resolution and unveiled a higher splicing efficiency of PB-associated XBP1 transcripts during unfolded protein response (UPR). Finally, based on single-cell LSM14A-TRIBE-ID (sc-LSM14A-TRIBE-ID), we demonstrated the dynamic pattern of mRNA-PB association during cell cycle progression.
Project description:Processing bodies (PBs) are dynamic, membraneless organelles consisting of RNAs and proteins. While PB proteins have been extensively characterized, the methods for systematically profiling PB-associated RNAs are limited. To address this, we developed PB-TRIBE-STAMP, a new tool based on two orthogonal RNA editing enzymes. Simultaneously applying APOBEC1-DDX6 and LSM14A-ADAR2dd, PB-TRIBE-STAMP identified 1,639 and 2,577 PB-associated mRNAs in HCT116 and HEK293T cells, respectively. Further genetic perturbation demonstrated that these transcripts were translationally repressed by PBs. Next, integration of PB-TRIBE-STAMP with long-read sequencing revealed that the PB-associated transcripts possessed shorter poly(A)-tails. Moreover, we established a TRIBE-ID-based tool to characterize the mRNA-PB association at high temporal resolution and unveiled a higher splicing efficiency of PB-associated XBP1 transcripts during unfolded protein response (UPR). Finally, based on sc-LSM14A-TRIBE-ID, we demonstrated the dynamic pattern of mRNA-PB interaction during cell cycle progression.
Project description:Most current methods to identify cell-specific RNA binding protein (RBP) targets require analyzing an extract, a strategy that is problematic with small amounts of material. We previously addressed this issue by developing TRIBE, a method that expresses an RBP of interest fused to the catalytic domain (cd) of the RNA editing enzyme ADAR. TRIBE performs Adenosine-to-Inosine editing on candidate RNA targets of the RBP. However, target identification is limited by the efficiency of the ADARcd. Here we describe HyperTRIBE, which carries a previously characterized hyperactive mutation (E488Q) of the ADARcd. HyperTRIBE identifies dramatically more editing sites than TRIBE, many of which are also edited by TRIBE but at a much lower editing frequency. The data have mechanistic implications for the enhanced editing activity of the HyperADARcd as part of a RBP fusion protein and also indicate that HyperTRIBE more faithfully recapitulates the known binding specificity of its RBP than TRIBE.