Enhancer trapping and annotation in Zebrafish mediated with Sleeping Beauty, piggyBac and Tol2 transposons

Although transposon-mediated enhancer trapping (ET) is successfully applied in diverse models, the efficiency of various transposon systems varies significantly, and little information is available regarding efficiency of enhancer trapping by various transposons in zebrafish. Most potential enhancers (Ens) still lack evidence of actual En activity. Here, we compared the differences in ET efficiency between sleeping beauty (SB), piggyBac (PB) and Tol2 transposons. Tol2 represented the highest germline transfer efficiencies at 55.56% (NF0 = 165), followed by SB (38.36%, NF0 = 151) and PB (32.65%, NF0 = 149). ET lines generated by the Tol2 transposon tended to produce offspring with a single expression pattern per line, while PB and SB tended to generate embryos with multiple expression patterns. In our tests, 10 putative Ens (En1⁻10) were identified by splinkerette PCR and comparative genomic analysis. Combining the GFP expression profiles and mRNA expression patterns revealed that En1 and En2 may be involved in regulation of the expression of dlx1a and dlx2a, while En6 may be involved in regulation of the expression of line TK4 transgene and rps26, and En7 may be involved in the regulation of the expression of wnt1 and wnt10b. Most identified Ens were found to be transcribed in zebrafish embryos, and their regulatory function may involve eRNAs

Elicitation of bioactive compounds production in in vitro cultures of Panax ginseng

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Eliška Syslová Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Thesis title: Elicitation of bioactive compounds production in in vitro cultures of Panax ginseng This diploma thesis is focused on the study of affecting the biosynthesis of secondary metabolites of Panax ginseng C. A. Meyer and is focused on increasing the production of biologically active polyacetylenes and ginsenosides in in vitro root cultures by various elicitors. The effect of the concentration of elicitors and growth regulators on the production of root biomass and secondary metabolites was researched. The carriers of the main pharmacological actions of Panax ginseng are ginsenosides and polyacetylenes. The root cultures were cultivated in the liquid medium in the dark at 24 ± 1řC. During the 27 day the sterile elicitation was done and a week later it was harvested. The samples were processed according to the optimized procedure, separately for UPLC analysis of ginsenosides and UPLC analysis of polyacetylenes as well. The presence of individual elicitors influenced the growth of biomass and the amount of ginsenosides and polyacetylenes in the samples

Additional file 1: Table S1. of The contribution of transposable elements to size variations between four teleost genomes

Numbers of families of DNA repeats from RepBase and the de novo repeats identified by TBLASTN. (PDF 91 kb

An Efficient Asymmetric Biomimetic Transamination of α‑Keto Esters to Chiral α‑Amino Esters

An efficient asymmetric biomimetic transamination of α-keto esters with quinine derivatives as chiral bases was described. A wide variety of α-amino esters containing various functional groups can be synthesized in high yield and enantioselectivity

Expansion sequencing: Spatially precise in situ transcriptomics in intact biological systems

Methods for highly multiplexed RNA imaging are limited in spatial resolution and thus in their ability to localize transcripts to nanoscale and subcellular compartments. We adapt expansion microscopy, which physically expands biological specimens, for long-read untargeted and targeted in situ RNA sequencing. We applied untargeted expansion sequencing (ExSeq) to the mouse brain, which yielded the readout of thousands of genes, including splice variants. Targeted ExSeq yielded nanoscale-resolution maps of RNAs throughout dendrites and spines in the neurons of the mouse hippocampus, revealing patterns across multiple cell types, layer-specific cell types across the mouse visual cortex, and the organization and position-dependent states of tumor and immune cells in a human metastatic breast cancer biopsy. Thus, ExSeq enables highly multiplexed mapping of RNAs from nanoscale to system scale