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A peptide-based silicon nanowire array sensor for the specific detection of TNT was developed, and the effect of the biointerfacing linker was investigated.

Functional genomics, synthetic biology and metabolic engineering require efficient tools to deliver long DNA fragments or multiple gene constructs. Although numerous DNA assembly methods exist, most are complicated, time-consuming and expensive. Here, we developed a simple and flexible strategy, unique nucleotide sequence-guided nicking endonuclease (UNiE)-mediated DNA assembly (UNiEDA), for efficient cloning of long DNAs and multigene stacking. In this system, a set of unique 15-nt 3' single-strand overhangs were designed and produced by nicking endonucleases (nickases) in vectors and insert sequences. We introduced UNiEDA into our modified Cre/loxP recombination-mediated TransGene Stacking II (TGSII) system to generate an improved multigene stacking system we call TGSII-UNiE. Using TGSII-UNiE, we achieved efficient cloning of long DNA fragments of different sizes and assembly of multiple gene cassettes. Finally, we engineered and validated the biosynthesis of betanin in wild tobacco (Nicotiana benthamiana) leaves and transgenic rice (Oryza sativa) using multigene stacking constructs based on TGSII-UNiE. In conclusion, UNiEDA is an efficient, convenient and low-cost method for DNA cloning and multigene stacking, and the TGSII-UNiE system has important application prospects for plant functional genomics, genetic engineering and synthetic biology research.

BackgroundWhile the contralateral seventh cervical nerve (CC7) cross transfer was designed to reconstruct paralyzed arm function after stroke, improvement in language function was found in patients combined with aphasia.ObjectiveTo evaluate the effect of improvement in language function after CC7 cross transfer in stroke patients with chronic aphasia and explore its potential mechanism.MethodsIn a prospective observative cohort, patients diagnosed with hemiplegia combined with aphasia were included. The language function was evaluated through the changes of Aphasia Quotient evaluated by Western Aphasia Battery (WAB-AQ) as well as its four subtests from baseline to 1 week and 6 months after the surgery. Patients also received oral agility test by Boston Diagnostic Aphasia Examination (BDAE-OA). Resting-state functional MRI (rs-fMRI) was scanned before and over 6 months after the surgery to explore the potential central mechanism in language improvements.ResultsThe average increase of WAB-AQ was 8.08 points from baseline to 1 week post-operatively (PConclusionsImmediate and stable improvement in language function was detected after CC7 cross transfer in hemiplegic patients combined with aphasia, which may be realized through enhanced function of language network in the bilateral hemisphere.

Supplementary Figure S1: Treatment with octyl-D-2-HG-induces MIR148A methylation; Supplementary Figure S2: Octyl-D-2-HG-induced MIR148A methylation status remains constant at late passages; Supplementary Figure S3: Octyl-D-2-HG induces CpG island methylation of the RBP1 promoter in 293T cells; Supplementary Figure S4: Octyl-D-2-HG induces CpG island methylation of the MIR148A promoter in normal human astrocytes (NHA); Supplementary Figure S5: MIR148A promoter CpG methylation in IDH1MUT-transfected 293T cells; Supplementary Figure S6: C227 prevents IDH1 mutant-induced MIR148A promoter methylation; Supplementary Figure S7: C227 prevents IDH1 mutant-induced RBP1 promoter methylation; Supplementary Figure S8: Schematic graph of MIR148A genomic location, CpG island, and promoter-reporter construct; Supplementary Figure S9: MIR148a expression is unchanged by octyl-D-2-HG treatment in early passages in 293T and NHA cells; Supplementary Figure S10: Octyl-D-2-HG withdrawal restores MIR148A promoter to an unmethylated state

Supplementary Materials and Methods

Supplementary Figures S1-10. Figure S1. RRBS methylation results of full miR-148a-associated CpG island. Figure S2. The methylation status of IDH1 mutation is prognostic in GBM and Anaplastic Astrocytoma (Grade III). Figure S3. miR-148a promotor CpG island was demethylated and expression was upregulated after demethylating agent 5-aza-CdR treatment. Figure S4. Knockdown of DNMT1 in U251 cells by using siRNA. Figure S5. Introduction of mutant IDH1 into HEK293T. Figure S6. Overexpression of miR-148a in 293T-IDH1MUT doesn't block the methylation of miR-148a and RBP1. Figure S7. miR-148a expression level in miR-148a stably transfected U251 and T98G cells. Figure S8. Cell cycle analysis of T98G-miR-148a stable cells. Figure S9. The expression of G-CIMP genes DLC1 and CIDEB are downregulated in IDH1MUT GBM patient tissue samples and glioma cell lines. Figure S10. Model summarizing the role of miR-148a in IDH1 mutant gliomas.