1,888 research outputs found
Efficient induction of heritable inversions in plant genomes using the CRISPR/Cas system
During the evolution of plant genomes, sequence inversions occurred repeatedly making the respective regions inaccessible for meiotic recombination and thus for breeding. Therefore, it is important to develop technologies that allow the induction of inversions within chromosomes in a directed and efficient manner. Using the Cas9 nuclease from Staphylococcus aureus (Sa Cas9), we were able to obtain scarless heritable inversions with high efficiency in the model plant Arabidopsis thaliana . Via deep sequencing, we defined the patterns of junction formation in wildâtype and in the nonâhomologous endâjoining (NHEJ ) mutant ku70â1 . Surprisingly, in plants deficient of KU 70, inversion induction is enhanced, indicating that KU 70 is required for tethering the local broken ends together during repair. However, in contrast to wildâtype, most junctions are formed by microhomologyâmediated NHEJ and thus are imperfect with mainly deletions, making this approach unsuitable for practical applications. Using eggâcellâspecific expression of Cas9, we were able to induce heritable inversions at different genomic loci and at intervals between 3 and 18â
kb, in the percentage range, in the T1 generation. By screening individual lines, inversion frequencies of up to the 10% range were found in T2. Most of these inversions had scarless junctions and were without any sequence change within the inverted region, making the technology attractive for use in crop plants. Applying our approach, it should be possible to reverse natural inversions and induce artificial ones to break or fix linkages between traits at will
Clinical associations between gout and multiple sclerosis, Parkinsonâs disease and motor neuron disease: record-linkage studies
Detection of cannabinoid receptor type 2 in native cells and zebrafish with a highly potent, cell-permeable fluorescent probe.
Despite its essential role in the (patho)physiology of several diseases, CB2R tissue expression profiles and signaling mechanisms are not yet fully understood. We report the development of a highly potent, fluorescent CB2R agonist probe employing structure-based reverse design. It commences with a highly potent, preclinically validated ligand, which is conjugated to a silicon-rhodamine fluorophore, enabling cell permeability. The probe is the first to preserve interspecies affinity and selectivity for both mouse and human CB2R. Extensive cross-validation (FACS, TR-FRET and confocal microscopy) set the stage for CB2R detection in endogenously expressing living cells along with zebrafish larvae. Together, these findings will benefit clinical translatability of CB2R based drugs
Cannabinoids and Dementia: A Review of Clinical and Preclinical Data
The endocannabinoid system has been shown to be associated with neurodegenerative diseases and dementia. We review the preclinical and clinical data on cannabinoids and four neurodegenerative diseases: Alzheimerâs disease (AD), Huntingtonâs disease (HD), Parkinsonâs disease (PD) and vascular dementia (VD). Numerous studies have demonstrated an involvement of the cannabinoid system in neurotransmission, neuropathology and neurobiology of dementias. In addition, several candidate compounds have demonstrated efficacy in vitro. However, some of the substances produced inconclusive results in vivo. Therefore, only few trials have aimed to replicate the effects seen in animal studies in patients. Indeed, the literature on cannabinoid administration in patients is scarce. While preclinical findings suggest causal treatment strategies involving cannabinoids, clinical trials have only assessed the suitability of cannabinoid receptor agonists, antagonists and cannabidiol for the symptomatic treatment of dementia. Further research is needed, including in vivo models of dementia and human studies
Pharmacologic inhibition of S-nitrosoglutathione reductase protects against experimental asthma in BALB/c mice through attenuation of both bronchoconstriction and inflammation
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