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Biology subjects

Ferry, Q. R.

Publications and source records attributed to Ferry, Q. R..

3 recordsLinked to original sources

LOAM: A family of genomic language models trained on long-read soil metagenomes

Soil ecosystems represent a vast, largely uncharacterised reservoir of microbial diversity. Metagenomic assembly has unlocked access to this resource, and recent advances in long-read sequencing have improved the recovery and quality of microbial genomes from complex samples. While genomic language models have proven highly effective at capturing biological concepts from large sequence datasets, they have predominantly been trained on reference genomes or short-read assemblies. Here, we present LOAM, a family of decoder-only genomic language models ranging from 25 to 624 million parameters and trained exclusively on Oxford Nanopore long-read environmental metagenomes. LOAM model performance scales predictably with model size and training-token budget. Despite a relatively small training sequence corpus, LOAM models outperformed comparably sized models across biological benchmarks, and achieved performance competitive with substantially larger state-of-the-art models trained on much larger datasets. Context-intervention experiments further showed that LOAM models use genomic information over several kilobases, highlighting the potential value of increased contiguity provided by long-read metagenome-assembled genomes. For probe-based benchmark tasks, we systematically evaluated representations across hidden layers and revealed that task-relevant biological information was frequently more linearly accessible from intermediate than final model layers. Finally, we observed that variation in zero-shot variant-effect prediction was strongly associated with the presence of homologous target sequences in the pre-training corpus. Together, these results establish long-read environmental metagenomes as a viable foundation for training competitive genomic language models and demonstrate the importance of both model scale and training-corpus composition for biological generalisation.

genomics↗

Specific Modulation of CRISPR Transcriptional Activators through RNA-Sensing Guide RNAs in Mammalian Cells and Zebrafish Embryos

Cellular transcripts encode important information regarding cell identity and disease status. The activation of CRISPR in response to RNA biomarkers holds the potential for controlling CRISPR activity with spatiotemporal precision. This would enable the restriction of CRISPR activity to specific cell types expressing RNA biomarkers of interest while preventing unwanted activity in other cells. Here, we present a simple and specific platform for modulating CRISPR activity in response to RNA detection through engineering Streptococcus pyogenes Cas9 single-guide RNAs (sgRNAs). sgRNAs are engineered to fold into complex secondary structures that, in the ground state, inhibit their activity. The engineered sgRNAs become activated upon recognising complementary RNAs, thus enabling Cas9 to perform its function. Our approach enables CRISPR activation in response to RNA detection in both HEK293T cells and zebrafish embryos. Iterative design optimisations allowed the development of computational tools for generating sgRNAs capable of detecting RNA sequences of choice. Mechanistic investigations reveal that engineered sgRNAs are cleaved during RNA detection, and we identify key positions that benefit from chemical modifications to improve the stability of engineered sgRNAs in vivo. Our sensors open up novel opportunities for developing new research and therapeutic applications using CRISPR activation in response to endogenous RNA biomarkers.

synthetic biology↗

Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression

Spatial/temporal control of Cas9 guide RNA expression could considerably expand the utility of CRISPR-based technologies. Current approaches based on tRNA processing offer a promising strategy but suffer from high background. Here we developed a variant screening platform to identify differential sequence determinants of human tRNA promoter and processing activities. Rational design based on the ensuing principles allowed us to engineer an improved tRNA scaffold that enabled highly specific guide RNA production from a Pol-II promoter.

bioengineering↗