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Palacios, S.

Publications and source records attributed to Palacios, S..

5 recordsLinked to original sources

Prediction of late blight severity in a large panel of potato genotypes using low-altitude aerial images and machine learning methods

Potato (Solanum tuberosum L.) is a staple crop crucial to global food security, yet its production is severely threatened by late blight (LB), caused by Phytophthora infestans, one of the most destructive plant diseases worldwide. Breeding programs for LB resistance have traditionally relied on labor-intensive and subjective visual assessments, which limit scalability and consistency, particularly in early-generation trials. Unmanned aerial vehicle (UAV)-based remote sensing combined with machine learning (ML) offers a promising alternative for objective, high-throughput disease phenotyping. This study evaluated the potential of UAV-derived multispectral imagery and ML techniques to estimate LB severity across large and genetically diverse potato breeding populations, comprising 2,745 clones in one trial and 492 accessions in another, conducted in Oxapampa, Pasco, Peru. We compared vegetation index-based approaches with a machine learning framework that integrates K-means clustering and Kernel Ridge Regression (KRR) and assessed their ability to capture genotypic variation and support selection decisions. NDVI consistently showed a strong correlation with visually assessed LB severity, particularly at advanced stages of disease development, enabling objective discrimination between healthy and diseased canopy tissues. However, the KRR-based approach outperformed linear NDVI-based models by capturing nonlinear relationships between spectral responses and disease progression. Estimates of LB severity derived from NDVI and KRR models, expressed as best linear unbiased estimates (BLUEs), showed strong and biologically consistent relationships with the area under the disease progress curve (AUDPC), particularly during later UAV acquisitions. Selection coincidence between UAV-derived estimates and AUDPC-based rankings was substantially higher at intermediate to advanced stages of disease progression, suggesting that UAV assessments at these stages may capture sufficient phenotypic variation to distinguish genotypes. These findings indicate that UAV-based multispectral phenotyping, especially when integrated with ML, provides a practical and scalable approach for assessing LB severity in potato breeding programs while reducing the need for time-consuming field evaluations.

plant biology↗

Reversing transgene silencing via targeted chromatin editing

Mammalian cell engineering offers the opportunity to uncover biological principles and develop next-generation biotechnologies. However, epigenetic silencing of transgenes hinders the control of gene expression in mammalian cells. Here, we use chromatin editing of an integrated reporter in CHO-K1 and human induced pluripotent stem cells to study the molecular interactions driving silencing and its reversal. After transient induction of either DNA methylation or H3K9me3, stable silencing was exclusively observed with both marks. Due to the positive feedback between DNA methylation and H3K9me3 and the relative low stability of H3K9me3, our model predicts that removing DNA methylation is sufficient for transgene reactivation. Accordingly, targeted DNA demethylation reactivated the reporter irrespective of whether silencing was achieved by inducing DNA methylation, H3K9me3, or by the endogenous cellular machinery. These results shed light on molecular mechanisms at play during silencing and provide engineering tools for potent and specific transgene reactivation in mammalian cells.

bioengineering↗

Analog epigenetic cell memory by graded DNA methylation

Chemical modifications to histones and DNA play a crucial role in the regulation of transcription and in the maintenance of chromatin states that are not permissive to gene expression [1-3]. However, the landscape of gene expression states that these modifications stably maintain remains uncharted. Here, we show that gene expression can be memorized at a wide range of levels thus implementing analog epigenetic memory. Mechanistically, we find that DNA methylation serves a primary role in maintaining memory across cell divisions while histone modifications only follow DNA methylation to regulate gene expression. Employing targeted epigenetic editing and time-course analysis, we analyzed the temporal stability of gene expression and DNA methylation post removal of epigenetic effectors. We found that the grade of DNA methylation in the genes promoter, defined as the mean fraction of methylated CpGs, remains stable over time and inversely correlates with gene expression level. By contrast, Histone 3 lysine 9 trimethylation (H3K9me3) could not persist after removal of its writer in the absence of DNA methylation. These experimental findings, combined with our chromatin modification model, indicate that the absence of positive feedback mechanisms around DNA methylation - unlike those found in histone modifications - enable the temporal stability of the DNA methylation grade, which leads to analog memory. These results expand current knowledge on how epigenetic memory is achieved in natural systems. Moreover, we anticipate that analog memory through graded DNA methylation will enable to program mammalian cells with fine-grained information storage. This capability will significantly enhance the sophistication of engineered cell functionality in applications including tissue engineering, organoids, and cell therapies.

bioengineering↗

Synthetic genetic circuits to uncover and enforce the OCT4 trajectories of successful reprogramming of human fibroblasts

Reprogramming human fibroblasts to induced pluripotent stem cells (iPSCs) is inefficient, with heterogeneity among transcription factor (TF) trajectories driving divergent cell states. Nevertheless, the impact of TF dynamics on reprogramming efficiency remains uncharted. Here, we identify the successful reprogramming trajectories of the core pluripotency TF, OCT4, and design a genetic controller that enforces such trajectories with high precision. By combining a genetic circuit that generates a wide range of OCT4 trajectories with live-cell imaging, we track OCT4 trajectories with clonal resolution and find that a distinct constant OCT4 trajectory is required for colony formation. We then develop a synthetic genetic circuit that yields a tight OCT4 distribution around the identified trajectory and outperforms in terms of reprogramming efficiency other circuits that less accurately regulate OCT4. Our synthetic biology approach is generalizable for identifying and enforcing TF dynamics for cell fate programming applications. One-sentence summaryGenetic controllers and live-cell imaging offer a versatile strategy for probing the role of transcription factor dynamics in cell fate transitions.

synthetic biology↗

PERSIST: A programmable RNA regulation platform using CRISPR endoRNases

Regulation of transgene expression is becoming an integral component of gene therapies, cell therapies and biomanufacturing. However, transcription factor-based regulation upon which the majority of such applications are based suffers from complications such as epigenetic silencing, which limits the longevity and reliability of these efforts. Genetically engineered mammalian cells used for cell therapies and biomanufacturing as well as newer RNA-based gene therapies would benefit from post-transcriptional methods of gene regulation, but few such platforms exist that enable sophisticated programming of cell behavior. Here we engineer the 5 and 3 untranslated regions of transcripts to enable robust and composable RNA-level regulation through transcript cleavage and, in particular, create modular RNA-level OFF- and ON-switch motifs. We show that genomically introduced transgenes exhibit resistance to silencing when regulated using this platform compared to those that are transcriptionally-regulated. We adapt nine CRISPR-specific endoRNases as RNA-level "activators" and "repressors" and show that these can be easily layered and composed to reconstruct genetic programming topologies previously achieved with transcription factor-based regulation including cascades, all 16 two-input Boolean logic functions, positive feedback, a feed-forward loop and a putative bistable toggle switch. The orthogonal, modular and composable nature of this platform as well as the ease with which robust and predictable gene circuits are constructed holds promise for their application in gene and cell therapies.

synthetic biology↗