8 research outputs found
Identification of a stable major-effect QTL (Parth 2.1) controlling parthenocarpy in cucumber and associated candidate gene analysis via whole genome re-sequencing
A New Generation of Lineage Tracing Dynamically Records Cell Fate Choices
Reconstructing the development of lineage relationships and cell fate mapping has been a fundamental problem in biology. Using advanced molecular biology and single-cell RNA sequencing, we have profiled transcriptomes at the single-cell level and mapped cell fates during development. Recently, CRISPR/Cas9 barcode editing for large-scale lineage tracing has been used to reconstruct the pseudotime trajectory of cells and improve lineage tracing accuracy. This review presents the progress of the latest CbLT (CRISPR-based Lineage Tracing) and discusses the current limitations and potential technical pitfalls in their application and other emerging concepts
Loss of Dip2b leads to abnormal neural differentiation from mESCs
Abstract Background Disco-interacting protein 2 homolog B is a member of the Dip2 family encoded by the Dip2b gene. Dip2b is widely expressed in neuro-related tissues and is essential in axonal outgrowth during embryogenesis. Methods Dip2b knockout mouse embryonic stem cell line was established by CRISPR/Cas9 gene-editing technology. The commercial kits were utilized to detect cell cycle and growth rate. Flow cytometry, qRT-PCR, immunofluorescence, and RNA-seq were employed for phenotype and molecular mechanism assessment. Results Our results suggested that Dip2b is dispensable for the pluripotency maintenance of mESCs. Dip2b knockout could not alter the cell cycle and proliferation of mECSs, or the ability to differentiate into three germ layers in vitro. Furthermore, genes associated with axon guidance, channel activity, and synaptic membrane were significantly downregulated during neural differentiation upon Dip2b knockout. Conclusions Our results suggest that Dip2b plays an important role in neural differentiation, which will provide a valuable model for studying the exact mechanisms of Dip2b during neural differentiation
Generation of Dip2a homozygous knockout murine ES cell line IBMSe001-A-1 via CRISPR/Cas9 technology
Structure-Based Design and Screen of Novel Inhibitors for Class II 3-Hydroxy-3-methylglutaryl Coenzyme A Reductase from Streptococcus Pneumoniae
3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR)
is a primary
target in the current clinical treatment of hypercholesterolemia with
specific inhibitors of “statin” family. Statins are
excellent inhibitors of the class I (human) enzyme but relatively
poor inhibitors of the class II enzyme, which are well-known as a
potential target to discover drugs fighting against the invasive diseases
originated from S. pneumoniae. However,
no significantly effective inhibitors of class II HMGR have been reported
so far. In the present study, the reasonable three-dimensional (3D)
structure of class II HMGR from S. pneumoniae (SP-HMGR-II) was built by Swissmodel. On the basis of the modeling
3D structure in “close” flap domain form, several novel
potential hit compounds out of SPECs database were picked out by using
structure-based screening strategy. Especially the compounds <b>4</b>, <b>3</b>, and <b>11</b> exhibit highly inhibitory
activities, with IC<sub>50</sub> values of 11.5, 18.5, and 18.1 ÎĽM,
respectively. Furthermore, the hit compounds were chosen as probe
molecules, and their probable interactions with the corresponding
individual residues have been examined by jointly using the molecular
docking, site-directed mutagenesis, enzymatic assays, and fluorescence
spectra, to provide an insight into a new special binding-model located
between the HMG-CoA and NADPH pockets. The good agreement between
theoretical and experimental results indicate that the modeling strategies
and screening processes in the present study are very likely to be
a promising way to search novel lead compounds with both structural
diversity and high inhibitory activity against SP-HMGR-II in the future