Generation of tooth profile for roots rotor based on virtual linkage associated with Assur group

This article, for the first time, presents the generation of Roots rotor tooth profiles based on an Assur-group-associated virtual linkage method. Taking the original Roots rotor as an example, structure and geometry of the Roots rotor are introduced, and based on the principle of inversion, an equivalent virtual linkage is identified for generating dedendum tooth profile of the rotor. Using linkage decomposition associated with elemental Assur groups, algorithm for computing the tooth curve is constructed leading to the explicit expression of rotor profile and the corresponding numerical simulation, verifying the validity of the proposed approach. For demonstration purpose, the virtual linkage method is then extended to the generation of tooth profiles for the variants of Roots rotors with arc-cycloidal curves and arc-involute curves. Integrated with computer-aided design, computer-aided engineering and computer-aided manufacturing software platforms, as well as the three-dimensional printing technology, this article provides an efficient and intuitive approach for Roots rotor system design, analysis and development

Efficient and versatile CRISPR engineering of human neurons in culture to model neurological disorders

The recent identification of multiple new genetic causes of neurological disorders highlights the need for model systems that give experimental access to the underlying biology. In particular, the ability to couple disease-causing mutations with human neuronal differentiation systems would be beneficial. Gene targeting is a well-known approach for dissecting gene function, but low rates of homologous recombination in somatic cells (including neuronal cells) have traditionally impeded the development of robust cellular models of neurological disorders. Recently, however, CRISPR/Cas9 gene editing technologies have expanded the number of systems within which gene targeting is possible. Here we adopt as a model system LUHMES cells, a commercially available diploid human female mesencephalic cell line that differentiates into homogeneous mature neurons in 1-2 weeks. We describe optimised methods for transfection and selection of neuronal progenitor cells carrying targeted genomic alterations using CRISPR/Cas9 technology. By targeting the endogenous X-linked MECP2 locus, we introduced four independent missense mutations that cause the autism spectrum disorder Rett syndrome and observed the desired genetic structure in 3-26% of selected clones, including gene targeting of the inactive X chromosome. Similar efficiencies were achieved by introducing neurodevelopmental disorder-causing mutations at the autosomal EEF1A2 locus on chromosome 20. Our results indicate that efficiency of genetic “knock-in” is determined by the location of the mutation within the donor DNA molecule. Furthermore, we successfully introduced an mCherry tag at the MECP2 locus to yield a fusion protein, demonstrating that larger insertions are also straightforward in this system. We suggest that our optimised methods for altering the genome of LUHMES cells make them an attractive model for the study of neurogenetic disorders