Nucleic Acids Research最新文献_第10页

DragonRNA: Generality of DNA-primed RNA-extension activities by DNA-directed RNA polymerases.

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf236

Emily Greenwald, Drew Galls, Joon Park, Nimit Jain, Stephen B Montgomery, Bijoyita Roy, Y Whitney Yin, Andrew Z Fire

{"title":"DragonRNA: Generality of DNA-primed RNA-extension activities by DNA-directed RNA polymerases.","authors":"Emily Greenwald, Drew Galls, Joon Park, Nimit Jain, Stephen B Montgomery, Bijoyita Roy, Y Whitney Yin, Andrew Z Fire","doi":"10.1093/nar/gkaf236","DOIUrl":"10.1093/nar/gkaf236","url":null,"abstract":"RNA polymerases (RNAPs) transcribe DNA into RNA. Several RNAPs, including from bacteriophages Sp6 and T7, Escherichia coli, and wheat germ, had been shown to add ribonucleotides to DNA 3' ends. Mitochondria have their own RNAPs (mtRNAPs). Examining reaction products of RNAPs acting on DNA molecules with free 3' ends, we found yeast and human mtRNAP preparations exhibit a robust activity of extending DNA 3' ends with ribonucleotides. The resulting molecules are serial DNA→RNA chains with the input DNA on the 5' end and extended RNA on the 3' end. Such chains were produced from a wide variety of DNA oligonucleotide inputs with short complementarity in the sequence to the DNA 3' end with the sequence of the RNA portion complementary to the input DNA. We provide a set of fluorescence-based assays for facile detection of such products and show that this activity is a general property of diverse RNAPs, including phage RNAPs and multi-subunit E. coli RNAP. These results support a model in which DNA serves as both primer and template, with extension beginning when the 3' end of the DNA is elongated with a ribonucleotide. As this DNA→RNA class of molecule remains unnamed, we propose the name DragonRNA.","PeriodicalId":19471,"journal":{"name":"Nucleic Acids Research","volume":"53 6","pages":""},"PeriodicalIF":16.6,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803930","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

DGCR8 haploinsufficiency leads to primate-specific RNA dysregulation and pluripotency defects.

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf197

Ana Colomer-Boronat, Lisanne I Knol, Guillermo Peris, Laura Sanchez, Silvia Peluso, Pablo Tristan-Ramos, Ana Gazquez-Gutierrez, Priscilla Chin, Katrina Gordon, Guillermo Barturen, Robert E Hill, Francisco J Sanchez-Luque, Jose Luis Garcia-Perez, Alasdair Ivens, Sara Macias, Sara R Heras

{"title":"DGCR8 haploinsufficiency leads to primate-specific RNA dysregulation and pluripotency defects.","authors":"Ana Colomer-Boronat, Lisanne I Knol, Guillermo Peris, Laura Sanchez, Silvia Peluso, Pablo Tristan-Ramos, Ana Gazquez-Gutierrez, Priscilla Chin, Katrina Gordon, Guillermo Barturen, Robert E Hill, Francisco J Sanchez-Luque, Jose Luis Garcia-Perez, Alasdair Ivens, Sara Macias, Sara R Heras","doi":"10.1093/nar/gkaf197","DOIUrl":"10.1093/nar/gkaf197","url":null,"abstract":"The 22q11.2 deletion syndrome (22qDS) is a human disorder where the majority of clinical manifestations originate during embryonic development. 22qDS is caused by a microdeletion in one chromosome 22, including DGCR8, an essential gene for microRNA (miRNA) production. However, the impact of DGCR8 hemizygosity on human development is still unclear. In this study, we generated two human pluripotent cell models containing a single functional DGCR8 allele to elucidate its role in early development. DGCR8+/- human embryonic stem cells (hESCs) showed increased apoptosis as well as self-renewal and differentiation defects in both the naïve and primed states. The expression of primate-specific miRNAs was largely affected, due to impaired miRNA processing and chromatin accessibility. DGCR8+/- hESCs also displayed a pronounced reduction in human endogenous retrovirus class H (HERVH) expression, a primate-specific retroelement essential for pluripotency maintenance. The reintroduction of miRNAs belonging to the primate-specific C19MC cluster as well as the miR-371-3 cluster rescued the defects of DGCR8+/- cells. Mechanistically, downregulation of HERVH by depletion of primate-specific miRNAs was mediated by KLF4. Altogether, we show that DGCR8 is haploinsufficient in humans and that miRNAs and transposable elements may have co-evolved in primates as part of an essential regulatory network to maintain stem cell identity.","PeriodicalId":19471,"journal":{"name":"Nucleic Acids Research","volume":"53 6","pages":""},"PeriodicalIF":16.6,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11941479/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143730840","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Structure and cooperative formation of a FLI1 filament on contiguous GGAA DNA sites.

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf205

Caixia Hou, Oleg V Tsodikov

引用次数: 0

Post-replicative lesion processing limits DNA damage-induced mutagenesis.

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf198

Katarzyna H Masłowska, Ronald P Wong, Helle D Ulrich, Vincent Pagès

{"title":"Post-replicative lesion processing limits DNA damage-induced mutagenesis.","authors":"Katarzyna H Masłowska, Ronald P Wong, Helle D Ulrich, Vincent Pagès","doi":"10.1093/nar/gkaf198","DOIUrl":"10.1093/nar/gkaf198","url":null,"abstract":"DNA lesions are a threat to genome stability. To cope with them during DNA replication, cells have evolved lesion bypass mechanisms: Translesion Synthesis (TLS), which allows the cell to insert a nucleotide directly opposite the lesion, with the risk of introducing a mutation, and error-free damage avoidance (DA), which uses homologous recombination to retrieve the genetic information from the sister chromatid. In this study, we investigate the timing of lesion bypass in yeast and its implications for the accuracy of the process. Our findings reveal that DNA polymerase η can bypass common, UV-induced cyclobutane pyrimidine dimers at the fork, immediately after encountering the blocking lesion. In contrast, TLS at (6-4) photoproducts and bulky G-AAF adducts, mediated by Rev1 and Pol ζ, takes place behind the fork, at post-replicative gaps that are generated downstream of the lesion after repriming. We show that in this latter situation, TLS competes with the DA pathway, thus reducing overall mutagenicity of damage bypass. Additionally, our study demonstrates that Exo1 nuclease influences the balance between TLS and DA by modulating the size of the post-replicative gaps.","PeriodicalId":19471,"journal":{"name":"Nucleic Acids Research","volume":"53 6","pages":""},"PeriodicalIF":16.6,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11925729/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143670490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Direct RNA sequencing of the Escherichia coli epitranscriptome uncovers alterations under heat stress. 大肠杆菌表转录组的直接 RNA 测序揭示了热胁迫下的变化。

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf175

Sebastián Riquelme-Barrios, Leonardo Vásquez-Camus, Siobhan A Cusack, Korinna Burdack, Dimitar Plamenov Petrov, G Nur Yeşiltaç-Tosun, Stefanie Kaiser, Pascal Giehr, Kirsten Jung

{"title":"Direct RNA sequencing of the Escherichia coli epitranscriptome uncovers alterations under heat stress.","authors":"Sebastián Riquelme-Barrios, Leonardo Vásquez-Camus, Siobhan A Cusack, Korinna Burdack, Dimitar Plamenov Petrov, G Nur Yeşiltaç-Tosun, Stefanie Kaiser, Pascal Giehr, Kirsten Jung","doi":"10.1093/nar/gkaf175","DOIUrl":"10.1093/nar/gkaf175","url":null,"abstract":"Modifications of RNA, known as the epitranscriptome, affect gene expression, translation, and splicing in eukaryotes, with implications for developmental processes, cancer, and viral infections. In prokaryotes, regulation at the level of the epitranscriptome is still poorly understood. Here, we used nanopore direct RNA sequencing of Escherichia coli to study RNA modifications and their changes under heat stress. With a single sequencing reaction, we detected most known modification types in ribosomal RNA (rRNA), transfer RNA (tRNA), and messenger RNA (mRNA). RNA sequencing was complemented by a multifaceted approach that included mass spectrometry, deletion mutants, single-nucleotide polymerase chain reaction, and in vitro methylation. Known 5-methylcytidine (m5C) and N6-methyladenosine (m6A) sites in the rRNA were confirmed, but these types of modifications could not be localized in the mRNA. In response to heat stress, levels of m5C, m6A, and N6,N6-dimethyladenosine increased in the 16S rRNA. Sequencing and mass spectrometry data demonstrated a decrease in tRNA modification abundance in the anticodon loop at 45°C. In general, mRNA modifications at 37°C were enriched in the coding regions of genes associated with general metabolism and RNA processing, which shifted to genes involved in cell wall synthesis and membrane transport under heat stress. This study provides new insights into the complexity of post-transcriptional regulation in bacteria.","PeriodicalId":19471,"journal":{"name":"Nucleic Acids Research","volume":"53 6","pages":""},"PeriodicalIF":16.6,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11925731/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143670482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Single-cell genome and transcriptome sequencing without upfront whole-genome amplification reveals cell state plasticity of melanoma subclones.

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf173

Koen Theunis, Sebastiaan Vanuytven, Irene Claes, Jarne Geurts, Florian Rambow, Daniel Brown, Michiel Van Der Haegen, Oskar Marin-Bejar, Aljosja Rogiers, Nina Van Raemdonck, Eleonora Leucci, Jonas Demeulemeester, Alejandro Sifrim, Jean-Christophe Marine, Thierry Voet

{"title":"Single-cell genome and transcriptome sequencing without upfront whole-genome amplification reveals cell state plasticity of melanoma subclones.","authors":"Koen Theunis, Sebastiaan Vanuytven, Irene Claes, Jarne Geurts, Florian Rambow, Daniel Brown, Michiel Van Der Haegen, Oskar Marin-Bejar, Aljosja Rogiers, Nina Van Raemdonck, Eleonora Leucci, Jonas Demeulemeester, Alejandro Sifrim, Jean-Christophe Marine, Thierry Voet","doi":"10.1093/nar/gkaf173","DOIUrl":"10.1093/nar/gkaf173","url":null,"abstract":"Single-cell multi-omics methods enable the study of cell state diversity, which is largely determined by the interplay of the genome, epigenome, and transcriptome. Here, we describe Gtag&T-seq, a genome-and-transcriptome sequencing (G&T-seq) protocol of the same single cells that omits whole-genome amplification (WGA) by using direct genomic tagmentation (Gtag). Gtag drastically decreases the cost and improves coverage uniformity at single-cell and pseudo-bulk levels compared to WGA-based G&T-seq. We also show that transcriptome-based DNA copy number inference has limited resolution and accuracy, underlining the importance of affordable multi-omic approaches. Applying Gtag&T-seq to a melanoma xenograft model before treatment and at minimal residual disease revealed differential cell state plasticity and treatment response between cancer subclones. In summary, Gtag&T-seq is a low-cost and accurate single-cell multi-omics method that explores genetic alterations and their functional consequences in single cells at scale.","PeriodicalId":19471,"journal":{"name":"Nucleic Acids Research","volume":"53 6","pages":""},"PeriodicalIF":16.6,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11941470/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143730849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

MAIGRET: a CRISPR-based immunoassay that employs antibody-induced cell-free transcription of CRISPR guide RNA strands.

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf238

Francesca Miceli, Sara Bracaglia, Daniela Sorrentino, Alessandro Porchetta, Simona Ranallo, Francesco Ricci

{"title":"MAIGRET: a CRISPR-based immunoassay that employs antibody-induced cell-free transcription of CRISPR guide RNA strands.","authors":"Francesca Miceli, Sara Bracaglia, Daniela Sorrentino, Alessandro Porchetta, Simona Ranallo, Francesco Ricci","doi":"10.1093/nar/gkaf238","DOIUrl":"10.1093/nar/gkaf238","url":null,"abstract":"Here we report on the development of a CRISPR-based assay for the sensitive and specific detection of antibodies and antigens directly in complex sample matrices. The assay, called Molecular Assay based on antibody-Induced Guide-RNA Enzymatic Transcription (MAIGRET), is based on the use of a responsive synthetic DNA template that triggers the cell-free in vitro transcription of a guide RNA strand upon recognition of a specific target antibody. Such transcribed guide RNA activates the DNA collateral activity of the Cas12a enzyme, leading to the downstream cleavage of a fluorophore/quencher-labeled reporter and thus resulting in an increase in the measured fluorescence signal. We have used MAIGRET for the detection of six different antibodies with high sensitivity (detection limit in the picomolar range) and specificity (no signal in the presence of non-target antibodies). MAIGRET can also be adapted to a competitive approach for the detection of specific antigens. With MAIGRET, we significantly expand the scope and applicability of CRISPR-based sensing approaches to potentially enable the measurement of any molecular target for which an antibody is available.","PeriodicalId":19471,"journal":{"name":"Nucleic Acids Research","volume":"53 6","pages":""},"PeriodicalIF":16.6,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11952961/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143743433","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

BioRels' data infrastructure: a scientific schema and exchange standard to transform and enhance biological data sciences.

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf254

Jibo Wang, Amanda Turney, Lauren Murray, Andrew M Craven, Patty Bragger-Wilkinson, Bruno Dos Santos, Jaroslav Martasek, Jeremy Desaphy

{"title":"BioRels' data infrastructure: a scientific schema and exchange standard to transform and enhance biological data sciences.","authors":"Jibo Wang, Amanda Turney, Lauren Murray, Andrew M Craven, Patty Bragger-Wilkinson, Bruno Dos Santos, Jaroslav Martasek, Jeremy Desaphy","doi":"10.1093/nar/gkaf254","DOIUrl":"10.1093/nar/gkaf254","url":null,"abstract":"Our understanding of biology and medicinal sciences augmented by advances in data structures and algorithms has resulted in proliferation of thousands of open-sourced resources, tools, and websites that are made by the scientific community to access, process, store, and visualize biological data. However, such data have become increasingly complex and heterogeneous, leading to an entangled web of relationships and external identifiers. Despite emergence of infrastructure such as data lakes, the scientists are still responsible for the time consuming and costly exercise to find, extract, clean, prepare, and maintain such data sources while following the FAIR principles. To better understand the complexity, we lay down a representation of the mainstream data ecosystem, describing the natural relationships and concepts found in biology. Built upon it and the fundamental principles of data unicity and atomicity, we introduce BioRels, an automated and standardized data preparation workstream aiming at improving reproducibility and speed for all scientists and handling up to 145 billion data points. BioRels allows complex querying capabilities across several data sources seamlessly and provides an exchange format, BIORJ, to export and import data with all its dependency and metadata. At last, we describe the advantages, limitations, applications, and perspectives of a future approach BioRels-KB to expand future data preparation capabilities.","PeriodicalId":19471,"journal":{"name":"Nucleic Acids Research","volume":"53 6","pages":""},"PeriodicalIF":16.6,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11969666/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143780778","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mechanism-based approach in designing patient-specific combination therapies for nonsense mutation diseases.

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf216

Saleem Y Bhat, Arpan Bhattacharya, Hong Li, Xiaonan Cui, John D Lueck, Yale E Goldman, Barry S Cooperman

{"title":"Mechanism-based approach in designing patient-specific combination therapies for nonsense mutation diseases.","authors":"Saleem Y Bhat, Arpan Bhattacharya, Hong Li, Xiaonan Cui, John D Lueck, Yale E Goldman, Barry S Cooperman","doi":"10.1093/nar/gkaf216","DOIUrl":"10.1093/nar/gkaf216","url":null,"abstract":"Premature termination codon (PTC) diseases account for ∼12% of all human disease mutations. Although there are no FDA approved treatments for increasing PTC readthrough, one readthrough inducing drug, ataluren, has conditional approval for treatment of Duchenne muscular dystrophy elsewhere. Ataluren displays low toxicity in clinical trials for treatment of PTC diseases, but its therapeutic effects are inconsistent. The messenger RNA (mRNA) sequence context of a PTC is a major determinant of PTC readthrough efficiency. We have shown that ataluren stimulates readthrough exclusively by competitively inhibiting release factor complex (RFC) catalysis of translation termination. Here, using an in vitro reconstituted system, we demonstrate that PTC identity and the immediately adjacent mRNA sequence contexts modulate RFC activity in terminating peptide elongation. Such modulation largely determines the effectiveness of ataluren in stimulating readthrough, whether added alone or in combination with either the aminoglycoside G418 or an anticodon edited aa-tRNA, which stimulate readthrough by mechanisms orthogonal to that of ataluren. Our results suggest a potential rationale for the variability of ataluren effectiveness in stimulating readthrough. We hypothesize that patients harboring a PTC mutation within a sequence context promoting strong interaction with RFC will be resistant to ataluren, but that ataluren treatment will be more effective for patient sequences conferring weaker interaction with RFC.","PeriodicalId":19471,"journal":{"name":"Nucleic Acids Research","volume":"53 6","pages":""},"PeriodicalIF":16.6,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11954524/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143743437","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants.

IF 16.6 2区生物学

Nucleic Acids Research Pub Date : 2025-03-20 DOI: 10.1093/nar/gkaf037

Carlos A Vasquez, Nicola R B Osgood, Marcanthony U Zepeda, Dominika K Sandel, Quinn T Cowan, Malalage N Peiris, Daniel J Donoghue, Alexis C Komor

{"title":"Precision genome editing and in-cell measurements of oxidative DNA damage repair enable functional and mechanistic characterization of cancer-associated MUTYH variants.","authors":"Carlos A Vasquez, Nicola R B Osgood, Marcanthony U Zepeda, Dominika K Sandel, Quinn T Cowan, Malalage N Peiris, Daniel J Donoghue, Alexis C Komor","doi":"10.1093/nar/gkaf037","DOIUrl":"10.1093/nar/gkaf037","url":null,"abstract":"Functional characterization of genetic variants has the potential to advance the field of precision medicine by enhancing the efficacy of current therapies and accelerating the development of new approaches to combat genetic diseases. MUTYH is a DNA repair enzyme that recognizes and repairs oxidatively damaged guanines [8-oxoguanine (8-oxoG)] mispaired with adenines (8-oxoG·A). While some mutations in the MUTYH gene are associated with colorectal cancer, most MUTYH variants identified in sequencing databases are classified as variants of uncertain significance. Convoluting clinical classification is the absence of data directly comparing homozygous versus heterozygous MUTYH mutations. In this study, we present the first effort to functionally characterize MUTYH variants using precision genome editing to generate heterozygous and homozygous isogenic cell lines. Using a MUTYH-specific lesion reporter in which we site-specifically incorporate an 8-oxoG·A lesion in a fluorescent protein gene, we measure endogenous MUTYH enzymatic activity and classify them as pathogenic or benign. Further, we modify this reporter to incorporate the MUTYH repair intermediate (8-oxoG across from an abasic site) and validate it with co-immunoprecipitation experiments to demonstrate its ability to characterize the mechanism by which MUTYH mutants are defective at DNA repair.","PeriodicalId":19471,"journal":{"name":"Nucleic Acids Research","volume":"53 6","pages":""},"PeriodicalIF":16.6,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11952967/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143743452","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0