Application and Development of CRISPR/Cas9 Technology in Pig Research

Pigs provide valuable meat sources, disease models, and research materials for humans. However, traditional methods no longer meet the developing needs of pig production. More recently, advanced biotechnologies such as SCNT and genome editing are enabling researchers to manipulate genomic DNA molecules. Such methods have greatly promoted the advancement of pig research. Three gene editing platforms including ZFNs, TALENs, and CRISPR/Cas are becoming increasingly prevalent in life science research, with CRISPR/Cas9 now being the most widely used. CRISPR/Cas9, a part of the defense mechanism against viral infection, was discovered in prokaryotes and has now developed as a powerful and effective genome editing tool that can introduce and enhance modifications to the eukaryotic genomes in a range of animals including insects, amphibians, fish, and mammals in a predictable manner. Given its excellent characteristics that are superior to other tailored endonucleases systems, CRISPR/Cas9 is suitable for conducting pig-related studies. In this review, we briefly discuss the historical perspectives of CRISPR/Cas9 technology and highlight the applications and developments for using CRISPR/Cas9-based methods in pig research. We will also review the choices for delivering genome editing elements and the merits and drawbacks of utilizing the CRISPR/Cas9 technology for pig research, as well as the future prospects

The Total Flavonoid Extract from Glycyrrhiza inflat Bat. Suppresses Atrophic Gastritis in Rats through the Akt/MAPK Pathway

Ethnopharmacological Relevance. Glycyrrhiza inflat Bat. is widely used to treat gastric ulcer and gastritis in clinic in China. Aim of the Study. To investigate the protective effects and possible mechanisms of the total flavonoid extract (TFE) from G. inflat Bat. on atrophic gastritis (AG) rats. Materials and Methods. The rat AG model was established by providing sodium deoxycholate and alcohol, and then, AG rats were treated with TFE for 30 days. Pathologic changes in gastric specimens were observed using hematoxylin and eosin staining, and the capability of gastric mucosa to secrete mucus was examined by alcian blue-periodic acid Schiff staining. Apoptosis induction in gastric tissues was measured by the TUNEL assay. The expressions of Bcl-2, Bax, and proteins in the Akt/MAPK pathway in gastric tissues were examined by immunohistochemistry and/or Western blotting. Results. Compared with the AG group, TFE attenuated the damage of gastric mucosa as reflected by the thickening of the lamina propria and the thinning of the muscularis mucosae. Moreover, TFE induced apoptosis in gastric mucosa with increasing Bax/Bcl-2 expression ratio. Concomitantly, the degrees of p-ErkThr202/Tyr204 and p-AktThr308 were decreased, whereas those of p-p38Thr180/Tyr182 and p-JNKThr183/Tyr185 were increased. Conclusion. We demonstrated the anti-AG effect of G. inflat Bat. in vivo and elucidated the underlying mechanisms that involve gastric mucosa protection through the Akt/MAPK pathway. The study provides a rationale for the application of G. inflat Bat. in the treatment of AG

Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P-associated external guide sequences

External guide sequences (EGSs) signify the short RNAs that induce ribonuclease P (RNase P), an enzyme responsible for processing the 5' termini of tRNA, to specifically cleave a target mRNA by forming a precursor tRNA-like complex. Hence, the EGS technology may serve as a potential strategy for gene-targeting therapy. Our previous studies have revealed that engineered EGS variants induced RNase P to efficiently hydrolyze target mRNAs. In the present research, an EGS variant was designed to be complementary to the mRNA coding for human cytomegalovirus (HCMV) major capsid protein (MCP), which is vital to form the viral capsid. In vitro, the EGS variant was about 80-fold more efficient in inducing human RNase P-mediated cleavage of the target mRNA than a natural tRNA-derived EGS. Moreover, the expressed variant and natural tRNA-originated EGSs led to a decrease of MCP expression by 98% and 73%-74% and a decrease of viral growth by about 10,000- and 200-fold in cells infected with HCMV, respectively. These results reveal direct evidence that the engineered EGS variant has higher efficiency in blocking the expression of HCMV genes and viral growth than the natural tRNA-originated EGS. Therefore, our findings imply that the EGS variant can be a potent candidate agent for the treatment of infections caused by HCMV

Additional file 1 of Gli1-mediated tumor cell-derived bFGF promotes tumor angiogenesis and pericyte coverage in non-small cell lung cancer

Supplementary Material 1