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z_;+j*Us#ezikH8Q&6Hj$d!W0Nn>L9*`4;S;s{==!>U4P)o%+Y02w*$t4XmPH`~6YP zohL#o^dAMOG5;3rf4_)2nUL&N_6~E~JJRmeXUQ9^1*D(eDGEZP&*!??I=HEqOpQAt* ztjGT582*1A@CWp}3)1v?|9QOse9#kxE|}>{K-Pb1;J+96|2*%|1^2zH#QAfaKOeH8 uj*!=hZ!iC-M*f^+cLeBy|9`kxkHQJ5TI2B~h^R2o9~ntSiE=T6!2boT{lrWF literal 0 HcmV?d00001 From d5ddff1fe3dab44a8c6eb694c7f1a4294c454d0e Mon Sep 17 00:00:00 2001 From: =?UTF-8?q?J=C4=99drzej=20Kubica?= Date: Fri, 1 Sep 2023 16:00:46 +0200 Subject: [PATCH 2/2] Update README.md --- README.md | 122 ++++++++++++++++++++++++++++++------------------------ 1 file changed, 69 insertions(+), 53 deletions(-) diff --git a/README.md b/README.md index 1545e5a..81f4572 100644 --- a/README.md +++ b/README.md @@ -4,33 +4,19 @@ ![ISOCOMP](https://i.ibb.co/vHLhrZq/Isocomp-logo1.png) -## Contributors -1. Yutong Qiu (Carnegie Mellon) -2. Chia Sin Liew (University of Nebraska-Lincoln) -3. Chase Mateusiak (Washington University) -4. Rupesh Kesharwani (Baylor College of Medicine) -5. Bida Gu (University of Southern California) -6. Muhammad Sohail Raza (Beijing Institute of Genomics, Chinese Academy of Sciences/China National Center for Bioinformation) -7. Evan Biederstedt (HMS) -8. Umran Yaman (UK Dementia Research Institute, University College London) -9. Abdullah Al Nahid (Shahjalal University of Science and Technology) -10. Trinh Tat (Houston Methodist Research Institute) -11. Sejal Modha (Theolytics Limited) -12. Jędrzej Kubica (University of Warsaw) - -## Github Codespace for Development - -To use codespaces for development purposes, do the following: - -1. fork the repo -2. switch to the 'develop' branch - - **NOTE**: if you plan to code/add a feature, create a branch from the 'develop' branch. Switch to it, and then continue on with the steps below. -4. click the green 'code' button. **But**, rather than copying the https or ssh link, click the tab that says "Codespace" -5. click the button that says "create codespace on develop". Go make some tea -- it takes ~5 minutes or so to set up the environment. But, once it is set up, you - will have a fully functioning vscode environment with all the dependencies installed. Start running the tests, set some breakpoints, take a look around! - -## Detailed project overview -https://github.com/collaborativebioinformatics/isocomp/blob/main/FinalPresentation_BCM_Hackathon_12Oct2022.pdf +## Contributors: +- Yutong Qiu (Carnegie Mellon) +- Chia Sin Liew (University of Nebraska-Lincoln) +- Chase Mateusiak (Washington University) +- Rupesh Kesharwani (Baylor College of Medicine) +- Bida Gu (University of Southern California) +- Muhammad Sohail Raza (Beijing Institute of Genomics, Chinese Academy of Sciences/China National Center for Bioinformation) +- Evan Biederstedt (HMS) +- Umran Yaman (UK Dementia Research Institute, University College London) +- Abdullah Al Nahid (Shahjalal University of Science and Technology) +- Trinh Tat (Houston Methodist Research Institute) +- Sejal Modha (Theolytics Limited) +- Jędrzej Kubica (University of Warsaw) ## Introduction NGS-targeted sequencing and WES have become routine for diagnosing Mendelian disease (Xue et al., 2015). Family sequencing (or "trio sequencing") involves sequencing a patient and parents (trio) or other relatives. This improves the diagnostic potential via the interpretation of germline mutations and enables the detection of de novo mutations which underlie most Mendelian disorders. @@ -47,10 +33,53 @@ This project aims to expand the applicability of long-read RNAseq for investigat -And what about gene fusions? We detect these in the stupidest possible way with short-read sequencing, and we think they're cancer-specific. What about the germline? +## Aim + +The aim of this project is to algorithmically characterize the "unique" (differing) isoforms between any number of samples using high-quality assembled isoforms. + +## Workflow +![](docs/images/isocomp_workflow.png) + +## Running the pipeline + +### Installation + +`pip install isocomp==0.3.0` + +For guidelines run: + +`isocomp --help` + +### Step 1. Create windows + +`isocomp create_windows -i sample1.gtf sample2.gtf sample3.gtf -f transcript -o clustered_file.gtf` + +### Step 2. Find unique isoforms across multiple samples + +`isocomp find_unique_isoforms -a clustered_file.gtf -f fasta_map.csv` + +File fasta_map.csv: + +``` +source,fasta +NA24385.filtered,BCM-data-HG002-All2Samples-hg38-Results/NA24385_HG002/MMSQANTI3Filter/NA24385.filtered.fasta +NA26105.filtered,BCM-data-HG002-All2Samples-hg38-Results/NA26105_GM26105/MMSQANTI3Filter/NA26105.filtered.fasta +``` + +### Example output + +For each isoform that is unique to at least one sample, we provide information about the read and the similarity between that isoform and the most similar isoform within the same window. + +The last column describes the normalized edit distance and the CIGAR string. -## Goals +``` +win_chr win_start win_end total_isoform isoform_name sample_from sample_compared_to mapped_start isoform_sequence selected_alignments +NC_060925.1 255178 288416 4 PB.6.2 HG004 HG002 255173 GGATTATCCGGAGCCAAGGTCCGCTCGGGTGAGTGCCCTCCGCTTTTT 0.02_HG002_PB.6.2_3=6I1=3I1286=11I +NC_060925.1 255178 288416 4 PB.6.2 HG004 HG005 255173 GGATTATCCGGAGCCAAGGTCCGCTCGGGTGAGTGCCCTCCGCTTTTTG 0.02_HG002_PB.6.2_3=6I1=3I1286=11 +``` -The goal of this project is to algorithmically characterize the "unique" (differing) isoforms between any number of samples using high-quality assembled isoforms. +## Detailed project overview +https://github.com/collaborativebioinformatics/isocomp/blob/main/FinalPresentation_BCM_Hackathon_12Oct2022.pdf ## Methods @@ -96,30 +125,6 @@ Isoseq3 (v3.2.2) generated HQ (Full-length high quality) transcripts [Table 1] w Differences between isoforms are categorized into [TODO] SNPs (<5bp), large-scale variants (>5bp), gene fusion, different exon usage, and completely novel sequences. These categories build upon those used by SQANTI to annotate disparities between sample isoforms and the reference transcriptome. Note that we extend the categories provided by SQANTI by adding SNPs and large-scale variants. -## Description - -## Flowchart -![](images/workflow.png) -### To extract sets of unique isoforms -![](images/workflow_part1.png) -### To annotate the unique isoforms -![](images/workflow_part2.png) - -## Example Output - -For each isoform that is unique to at least one sample, we provide information about the read and the similarity between that isoform and the most similar isoform within the same window. - -The last column describes the normalized edit distance and the CIGAR string. - -``` -win_chr win_start win_end total_isoform isoform_name sample_from sample_compared_to mapped_start isoform_sequence selected_alignments -NC_060925.1 255178 288416 4 PB.6.2 HG004 HG002 255173 GGATTATCCGGAGCCAAGGTCCGCTCGGGTGAGTGCCCTCCGCTTTTT 0.02_HG002_PB.6.2_3=6I1=3I1286=11I -NC_060925.1 255178 288416 4 PB.6.2 HG004 HG005 255173 GGATTATCCGGAGCCAAGGTCCGCTCGGGTGAGTGCCCTCCGCTTTTTG 0.02_HG002_PB.6.2_3=6I1=3I1286=11 -``` - -### Deployment - -Eventually, `pip install isocomp`. But not yet. ## DEPENDENCIES @@ -187,6 +192,17 @@ pip install poetry # and continue with the development install below ``` +## Github Codespace for Development + +To use codespaces for development purposes, do the following: + +1. fork the repo +2. switch to the 'develop' branch + - **NOTE**: if you plan to code/add a feature, create a branch from the 'develop' branch. Switch to it, and then continue on with the steps below. +4. click the green 'code' button. **But**, rather than copying the https or ssh link, click the tab that says "Codespace" +5. click the button that says "create codespace on develop". Go make some tea -- it takes ~5 minutes or so to set up the environment. But, once it is set up, you + will have a fully functioning vscode environment with all the dependencies installed. Start running the tests, set some breakpoints, take a look around! + ### Development Install [poetry](https://python-poetry.org/) and consider setting [the configuration