Background: The prevalence of Parkinson’s disease (PD), a chronic and progressive neurodegenerative disorder, is projected to increase twofold by 2030. Leucine-rich repeat kinase 2 (LRRK2) is the most commonly observ...Background: The prevalence of Parkinson’s disease (PD), a chronic and progressive neurodegenerative disorder, is projected to increase twofold by 2030. Leucine-rich repeat kinase 2 (LRRK2) is the most commonly observed gene in both familial and sporadic PD cases. Notably, there is a substantial augmentation in motor activity during both larval and adult stages of zebrafish lacking the lrrk2 gene. Nevertheless, the precise genetic abnormalities accountable for eliciting these phenotypes in zebrafish are yet to be elucidated. Methods: Real-time polymerase chain reaction (PCR) was conducted on zebrafish larvae at 6 days post fertilization (dpf) belonging to both the wild-type and lrk2(-/-) groups. Guide RNA was designed and subsequently employed in the PCR process. Electrophoresis was performed to facilitate identification. Results: The expression of CNTF mRNA was significantly diminished in lrrk2(-/-), in comparison to the wildtype zebrafish larvae. This finding implies that CNTF may have crucial implications in the regulated functioning of lrrk2, which is widely acknowledged as the predominant genetic factor contributing to hereditary PD. The primers for CNTF DNA were meticulously designed, and the electrophoresis results of the PCR product were subsequently presented. The wild type zebrafish embryos were meticulously prepared for micro-injection, and the resulting efficiency identification displayed the presence of the mutant PCR product, which exhibited the presence of several debris. Conclusions: The present study demonstrates the successful generation of CNTF mutant zebrafish using the CRISPR/Cas9 genome editing technique. Further investigations are necessary to deepen our understanding of the exogenous CNTF gene’s functionality.展开更多
Over the last decade,oncolytic virus(OV)therapy has shown its promising potential in tumor treatment.The fact that not every patient can benefit from it highlights the importance for defining biomarkers that help pred...Over the last decade,oncolytic virus(OV)therapy has shown its promising potential in tumor treatment.The fact that not every patient can benefit from it highlights the importance for defining biomarkers that help predict patients’responses.As particular self-amplifying biotherapeutics,the anti-tumor effects of OVs are highly dependent on the host factors for viral infection and replication.By using weighted gene co-expression network analysis(WGCNA),we found matrix remodeling associated 8(MXRA8)is positively correlated with the oncolysis induced by oncolytic virus M1(OVM).展开更多
文摘Background: The prevalence of Parkinson’s disease (PD), a chronic and progressive neurodegenerative disorder, is projected to increase twofold by 2030. Leucine-rich repeat kinase 2 (LRRK2) is the most commonly observed gene in both familial and sporadic PD cases. Notably, there is a substantial augmentation in motor activity during both larval and adult stages of zebrafish lacking the lrrk2 gene. Nevertheless, the precise genetic abnormalities accountable for eliciting these phenotypes in zebrafish are yet to be elucidated. Methods: Real-time polymerase chain reaction (PCR) was conducted on zebrafish larvae at 6 days post fertilization (dpf) belonging to both the wild-type and lrk2(-/-) groups. Guide RNA was designed and subsequently employed in the PCR process. Electrophoresis was performed to facilitate identification. Results: The expression of CNTF mRNA was significantly diminished in lrrk2(-/-), in comparison to the wildtype zebrafish larvae. This finding implies that CNTF may have crucial implications in the regulated functioning of lrrk2, which is widely acknowledged as the predominant genetic factor contributing to hereditary PD. The primers for CNTF DNA were meticulously designed, and the electrophoresis results of the PCR product were subsequently presented. The wild type zebrafish embryos were meticulously prepared for micro-injection, and the resulting efficiency identification displayed the presence of the mutant PCR product, which exhibited the presence of several debris. Conclusions: The present study demonstrates the successful generation of CNTF mutant zebrafish using the CRISPR/Cas9 genome editing technique. Further investigations are necessary to deepen our understanding of the exogenous CNTF gene’s functionality.
基金We acknowledge Dr.Zhe Liu of the Guangdong Provincial Center for Disease Control and Prevention for his assistance with cryo-EM.Miss.Liumei Wu and Miss.Jing Tang assisted with transfection assay and RNA-scope assay.This work was supported by the National Natural Science Foundation of China(No.81973342,82073883,81772675,81903652,and 81802536)the National Key R&D Program of China(2021YFA0909800)the Science and Technology Program of Guangzhou(No.201707020003).
文摘Over the last decade,oncolytic virus(OV)therapy has shown its promising potential in tumor treatment.The fact that not every patient can benefit from it highlights the importance for defining biomarkers that help predict patients’responses.As particular self-amplifying biotherapeutics,the anti-tumor effects of OVs are highly dependent on the host factors for viral infection and replication.By using weighted gene co-expression network analysis(WGCNA),we found matrix remodeling associated 8(MXRA8)is positively correlated with the oncolysis induced by oncolytic virus M1(OVM).
