Journal: Viruses

Article Title: Genomic Characterization and gE/gI-Deleted Strain Construction of Novel PRV Variants Isolated in Central China

doi: 10.3390/v15061237

Figure Lengend Snippet: Construction and identification of the mutant virus SX1911-ΔgE/gI. ( A ) Strategy for constructing SX1911-ΔgE/gI using the CRISPR/Cas9 and LoxP systems. Two sgRNAs were designed to guide Cas9 to delete the gE and gI genes, and GFP was used for both positive and negative screening of mutant virus production. ( B ) Identification of SX1911-ΔgE/gI via IFA and PCR targeting the gE gene. ( C ) Multistep growth curve of SX1911 and SX1911-ΔgE/gI in Vero cells. ( D ) Plaque sizes of SX1911 and SX1911-ΔgE/gI in Vero cells. Data are presented as the mean ± SD, and an asterisk indicates a significant difference between SX1911 and SX1911-ΔgE/gI. ***: p < 0.001.

Article Snippet: sgRNAs targeting the gE and gI genes were designed using an online CRISPR tool ( https://www.genscript.com/gRNA-design-tool.html , accessed on 15 May 2021).

Techniques: Mutagenesis, Virus, CRISPR

Journal: Cell Death & Disease

Article Title: Transcription coactivator Cited1 acts as an inducer of trophoblast-like state from mouse embryonic stem cells through the activation of BMP signaling

doi: 10.1038/s41419-018-0991-1

Figure Lengend Snippet: a Schematic representation of the strategy for Cited1 knockout (KO) by the CRISPR/Cas9 approach. The PAM sequences are in the rectangle. The cleavage site is pointed out by the scissor and arrow. Positions of the designed primers for genomic PCR are shown as arrows. The sequence of gRNA is shown in red color. b Four Cited1 KO E14T ESC cell lines with frame-shifted- or large fragment deleted- Cited1 were identified. This panel shows the genomic DNA PCR results. c Western blot analysis of protein levels of Cited1 in the parental wild-type (WT) ESCs and four Cited1 KO ESC lines after treatment with BMP4 for 6 days. d Morphology changes of parental wild-type (WT) ESCs and four Cited1 KO ESC lines before (top panel) and after (middle and bottom panels) treatment with BMP4 for 6 days. Scale bar: 200 μm (top and middle panels); 100 μm (bottom panel). e , f qRT-PCR analysis for expression levels of trophoblast ( e ) and pluripotency ( f ) markers in the four Cited1 KO ESC lines and their parental WT counterparts after treatment with BMP4 for 6 days. The comparison was made between Cited1 KO and WT ESCs at day 6 upon BMP4 treatment. The average mRNA level in untreated WT ESCs was set at 1.0. Data are shown as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The sgRNAs targeting Cited1 and Bmpr2 locus were designed using the CRISPR Design Tool ( http://crispr.mit.edu/ ) and synthesized by the Shanghai Shenggong Company.

Techniques: Knock-Out, CRISPR, Sequencing, Western Blot, Quantitative RT-PCR, Expressing, Comparison