查看更多>>摘要：Genetic lineage tracing has been widely employed to investigate cell lineages and fate.However,con-ventional reporting systems often label the entire cytoplasm,making it challenging to discern cell bound-aries.Additionally,single Cre-loxP recombination systems have limitations in tracing specific cell populations.This study proposes three reporting systems utilizing Cre,Dre,and Dre+Cre mediated recombination.These systems incorporate tdTomato expression on the cell membrane and PhiYFP expression within the nucleus,allowing for clear observation of the nucleus and membrane.The efficacy of these systems is successfully demonstrated by labeling cardiomyocytes and hepatocytes.The potential for dynamic visualization of the cell membrane is showcased using intravital imaging microscopy or three-dimensional imaging.Furthermore,by combining this dual recombinase system with the ProTracer sys-tem,hepatocyte proliferation is traced with enhanced precision.This reporting system holds significant importance for advancing the understanding of cell fate studies in development,homeostasis,and diseases.
查看更多>>摘要：Histone citrullination,an important post-translational modification mediated by peptidyl arginine deiminases,is essential for many physiological processes and epigenetic regulation.However,the causal relationship between histone citrullination and specific gene regulation remains unresolved.In this study,we develop a programmable epigenetic editor by fusing the peptidyl arginine deiminase(PAD)PPAD from Porphyromonas gingivalis with dCas9.With the assistance of gRNA,PPAD-dCas9 can recruit PPADs to specific genomic loci,enabling direct manipulation of the epigenetic landscape and regulation of gene expression.Our citrullination editor allows for the site-specific manipulation of histone H3R2,8,17 and H3R26 at target human gene loci,resulting in the activation or suppression of different genes in a locus-specific manner.Moreover,the epigenetic effects of the citrullination editor are specific and sustained.This epigenetic editor offers an ac-curate and efficient tool for exploring gene regulation of histone citrullination.
查看更多>>摘要：Multi-nucleotide variants(MNVs)are critical genetic variants associated with various genetic diseases.However,tools for precisely installing MNVs are limited.In this study,we present the development of a dual-base editor,BDBE,by integrating TadA-dual and engineered human N-methylpurine DNA glycosylase(eMPG)into nCas9(D10A).Our results demonstrate that BDBE effectively converts A-to-G/C/T(referred to as A-to-B)and C-to-T/G/A(referred to as C-to-D)simultaneously,yielding nine types of dinucleotides from adjacent CA nucleotides while maintaining minimal off-target effects.Notably,BDBE4 exhibits exceptional performance across multiple human cell lines and successfully simulated all nine dinucleotide MNVs from the gnomAD database.These findings indicate that BDBE significantly expands the product range of base editors and offers a valuable resource for advancing MNV research.
NETL
NSTL
万方数据