bioRxiv Science⌕ Search

Biology subjects

Tartaglia, J.

Publications and source records attributed to Tartaglia, J..

3 recordsLinked to original sources

MOAflow: how re-design a pipeline with Nextflow streamlines data analysis

BackgroundAdvances in high-throughput DNA sequencing technologies have dramatically reduced the time and cost required to generate genomic data. As sequencing is no longer a limiting factor, increasing attention must be paid to optimizing the analyses of the large-scale datasets produced. Efficient processing of such data is essential to reduce computational time and operational costs. In this context, workflow management systems (WMSs) have become key instruments for orchestrating complex bioinformatic pipelines. Among these systems, Nextflow has emerged as one of the most widely adopted solutions in bioinformatics. MethodsTo improve scalability and computational efficiency, we employed Nextflow to re-design an already existing pipeline dedicated to the analysis of MNase-defined cistrome-Occupancy (MOA-seq) data. The re-engineering process focused on modularizing the workflow and integrating containerization technologies to ensure reproducibility and easier deployment across heterogeneous computing environments. ResultsThe resulting workflow, named MOAflow, represents a modernized and fully containerized pipeline for MOA-seq data analysis. With only Docker and Nextflow required, the pipeline guarantees high portability and reproducibility. The data of the original article was used to benchmark the new pipeline. Its outputs closely match those of the original study with minor variations. ConclusionsMOAflow demonstrates how the adoption of robust WMS can substantially enhance the performance and usability of pre-existing bioinformatic pipelines. By leveraging containerization and Nextflow, it ensures consistent results across platforms while minimizing setup complexity. This work highlights the value of modern WMS-driven approaches in meeting the computational demands.

bioinformatics↗

Discovery of widespread activating mutations in a compact RNA-guided endonuclease

TnpB is a diverse family of RNA-guided endonucleases associated with prokaryotic transposons. Due to their small size and putative evolutionary relationship to CRISPR-Cas12, TnpB enzymes hold significant potential for genome editing. However, most TnpBs lack robust gene editing activity, and unbiased profiling of mutational effects on editing activity has not been explored. Here, we mapped comprehensive sequence-function landscapes of a TnpB ribonucleoprotein and discovered many activating mutations in both the protein and RNA. One- and two-position RNA mutants outperform existing variants, highlighting the utility of systematic RNA scaffold mutagenesis. Leveraging the proteins mutational landscape, we identified enhanced TnpB variants from a combinatorial library of activating mutations. These variants enhanced editing in human cells, N. benthamiana, pepper, and rice, with up to a fifty-fold increase compared to wild-type TnpB. These findings highlight previously unknown elements critical for regulating TnpB endonuclease activity and reveal surprising latent activity accessible through mutation.

molecular biology↗

Boosting with ALVAC-HIV and AIDSVAX B/E enhances Env constant region 1 and 2 antibody-dependent cellular cytotoxicity

Induction of protective antibodies is a critical goal of HIV-1 vaccine development. One strategy is to induce non-neutralizing antibodies that kill virus-infected cells as these antibody specificities have been implicated in slowing HIV-1 disease progression and in protection. HIV-1 Env constant region 1 and 2 (C1C2) antibodies frequently contain potent antibody dependent cellular cytotoxicity (ADCC) making them a vaccine target. Here we explore the effect of delayed and repetitive boosting of RV144 vaccinee recipients with ALVAC/AIDSVAX B/E on the C1C2-specific antibody repertoire. It was found that boosting increased clonal lineage specific ADCC breadth and potency. A ligand crystal structure of a vaccine-induced broad and potent ADCC-mediating C1C2-specific antibody showed that it bound a highly conserved Env gp120 epitope. Thus, rationally designed boosting strategies to affinity mature these type of IgG C1C2-specific antibody responses may be one method by which to make an improved HIV vaccine with higher efficacy than seen in the RV144 trial.\n\nSignificanceOver one million people become infected with HIV-1 each year making the development of an efficacious HIV-1 vaccine an important unmet medical need. The RV144 human HIV-1 vaccine-regimen is the only HIV-1 clinical trial to date to demonstrate vaccine-efficacy. An area of focus has been on identifying ways by which to improve upon RV144 vaccine-efficacy. The RV305 HIV-1 vaccine-regimen was a follow-up boost of RV144 vaccine-recipients that occurred 6-8 years after the conclusion of RV144. Our studies focused on the effect of delayed boosting in humans on the vaccine-induced antibody repertoire. It was found that boosting with a HIV-1 Env vaccine increased antibody-mediated effector function potency and breadth.

immunology↗