#+startup: showall * http server ** guix #+begin_src shell guix shell -m manifest.scm -- make run #+end_src #+RESULTS: ** by hand start a python http server with: 1. to compile this org document, call ~org-babel-tangle~ (=C-c C-v t=), which will produce =index.html=. alternately, you can just call =make= (a =manifest.scm= is provided for guix) 2. start an http server quickly in python on port 8000 to work around cors issues with file uris: #+begin_src shell python3 -m http.server -b :: 8000 #+end_src 3. then navigate to http://localhost:8000/ * html skeleton #+name: skeleton #+begin_src html :noweb yes :tangle index.html functional annotation for varscan

functional annotation for varscan

#+end_src * javascript ** entry point set up the javascript code by loading direct module dependencies and running the analysis when the form is submitted. #+begin_src javascript :noweb yes :tangle main.mjs import process from './varscan.mjs'; import Log from './logging.mjs'; function init() { console.info('initializing'); const varscanForm = document.querySelector('#varscan'); varscanForm.onsubmit = (event) => { submitForm(varscanForm); event.preventDefault(); }; const variantsInput = varscanForm.querySelector('input[type="file"]'); variantsInput.onchange = (event) => { Log.info("file uploaded"); submitButton.disabled = variantsInput.value === ""; }; const submitButton = varscanForm.querySelector('button'); submitButton.disabled = variantsInput.value === ""; } init(); #+end_src *** form submission when the form is submitted, load the reference genome,its protein coding regions, the uploaded variant data, and spit out the changed proteins. #+name: submit-form #+begin_src javascript :noweb yes :tangle main.mjs function submitForm(form) { Log.clear(); <> <> <> const filename = form .querySelector('input[name="variants-data"]') .files[0] .name; const extIndex = filename.lastIndexOf('.'); const resultsFilename = filename.substring(0, extIndex) + "-varscan" + filename.substring(extIndex); Promise.all([variantsPromise, genomePromise, protein2PosPromise]) .then(([variants, genome, protein2Pos]) => { const results = process(genome, protein2Pos, variants); updateDownloadLink(resultsFilename, results); fillTable(results); }); } #+end_src *** TODO variants upload we don't need to upload the file, but we're going to pretend we do for ux reasons. this just returns a promise for the “uploaded” file's csv data. - [ ] verify file format is csv without (or with) headers #+name: upload-variant #+begin_src javascript const variantsPromise = new Promise((resolve, reject) => { const file = form.querySelector('input[name="variants-data"]').files[0]; const reader = new FileReader(); reader.onload = (event) => { resolve(event.target.result); }; reader.readAsText(file); }); #+end_src *** reference genome promise load the reference genome from the server based on the filename in the form. #+name: reference-genome #+begin_src javascript const referenceGenomeFile = form.querySelector('select[name="reference-genome"]').value; const genomePromise = fetch(referenceGenomeFile) .then((response) => response.text()) .then((text) => text.replace('\r', '').replace('\n', '')) .catch((err) => Log.error("couldn't load reference genome:", err)); #+end_src *** TODO protein coding regions promise load the protein coding regions from the server based on the filename in the form. - [ ] this should probably be tied to the reference genome #+name: protein-coding-region #+begin_src javascript const proteinCodingRegionsFile = form.querySelector('select[name="protein-coding-regions"]').value; const protein2PosPromise = fetch(proteinCodingRegionsFile) .then((response) => response.text()) .then((text) => { return text .replace('\r', '') .split('\n') .reduce( (acc, line) => { const [name, start, stop] = line.split(','); acc[name] = [Number(start), Number(stop)]; return acc; }, {}); }) .catch((err) => Log.error("couldn't load protein coding regions:", err)); #+end_src *** display results take the results and stuff them in the output table #+begin_src javascript :noweb yes :tangle main.mjs function fillTable(results) { const outputDiv = document.querySelector('#results') outputDiv.hidden = false; const tbody = outputDiv.querySelector('table tbody'); // clear the table while (tbody.firstChild) { tbody.removeChild(tbody.firstChild); } results.forEach((row) => { const tr = document.createElement('tr'); row.forEach((col) => { const td = document.createElement('td'); td.innerHTML = col; tr.appendChild(td); }) tbody.appendChild(tr); }); } #+end_src create a download link #+begin_src javascript :noweb yes :tangle main.mjs function updateDownloadLink(filename, results) { const anchor = document.querySelector('#download'); if (anchor.url) { window.URL.revokeObjectURL(anchor.url); anchor.url = undefined; } const data = results.map((row) => row.join(",") + "\r\n"); const blob = new Blob( data, { type: 'text/csv' }); const url = window.URL.createObjectURL(blob); anchor.href = url; anchor.download = filename; return anchor; } #+end_src ** variant to codon whatever i need to name this #+begin_src javascript :tangle varscan.mjs import Log from './logging.mjs'; import codon2AA from './codon2AminoAcid.mjs'; import aa2Code from './aminoAcid2Code.mjs'; function process(genome, protein2Pos, variants) { let lineno = 0; let name = 'unnamed-change'; return variants .split('\n') .reduce( (acc, line) => { lineno++; line.replace('\r', ''); if (line === "") { return acc; } const [nameMaybe, ref, posStr, origNucleotide, newNucleotide] = line.split(','); name = nameMaybe || name; const pos = Number(posStr); const [protein, proteinStart, aaIndex] = findProtein(pos, protein2Pos); if (!protein) { return acc.concat([[ name, origNucleotide.toLowerCase()+posStr+newNucleotide.toLowerCase(), '', "non-coding" ]]); } // start of codon relative to start of protein, 0-based const changedStart = pos - proteinStart; // convert position to start of the codon and the // offset of the change from the start of the codon. const offset = changedStart % 3; const codonStart = changedStart - offset; // pos points to the change within the genome (1-index) // proteinStart points to the start of the changed protein within the genome (1-index) // changedStart points to the changed nucleotide within the protein (0-index) // codonStart points to the start of the changed codon within the protein (0-index) // offset points to the changed nucleotide within the codon (0-index) // position of codon within entire genome (0-index) const absCodonStart = proteinStart + codonStart - 1; const origCodon = genome.substring(absCodonStart, absCodonStart + 3); const origAA = aa2Code[codon2AA[origCodon]]; const newCodon = origCodon.substring(0, offset) + newNucleotide + origCodon.substring(offset+1); const newAA = aa2Code[codon2AA[newCodon]]; // check the change against the reference genome. const checkNucleotide = origCodon[offset]; if (checkNucleotide !== origNucleotide) { const checkCodon = origCodon.substring(0, offset) + origNucleotide + origCodon.substring(offset+1); const checkAA = aa2Code[codon2AA[checkCodon]]; const aaChange = `${checkAA}${aaIndex}${newAA}` Log.warn(`${name} (line ${lineno}): nucleotide at position ${pos} is “${checkNucleotide.toLowerCase()}” in the reference, but “${origNucleotide.toLowerCase()}” was supplied. If the supplied nucleotide is correct, then the amino acid change is ${aaChange}.`) } const nucleotideChange = `${checkNucleotide.toLowerCase()}${pos}${newNucleotide.toLowerCase()}` const aaChange = `${origAA}${aaIndex}${newAA}` return acc.concat([[ name, nucleotideChange, protein, aaChange ]]); }, []); } // pos is 1-based index // // returns protein name, 1-index of start of protein in genome, and // 1-index of of offset of `pos` in its codon. function findProtein(pos, protein2Pos) { for (const name in protein2Pos) { const [start, end] = protein2Pos[name]; if (start <= pos && pos <= end) { // normal people count from 1, not 0 const index = Math.floor((pos - start) / 3) + 1; return [name, start, index]; } } return []; } export default process #+end_src ** codon to amino acid table :PROPERTIES: :VISIBILITY: folded :END: #+begin_src javascript :noweb yes :tangle codon2AminoAcid.mjs const codon2AA = { 'GCT': 'Ala', 'GCC': 'Ala', 'GCA': 'Ala', 'GCG': 'Ala', 'CGT': 'Arg', 'CGC': 'Arg', 'CGA': 'Arg', 'CGG': 'Arg', 'AGA': 'Arg', 'AGG': 'Arg', 'AAT': 'Asn', 'AAC': 'Asn', 'GAT': 'Asp', 'GAC': 'Asp', 'TGT': 'Cys', 'TGC': 'Cys', 'CAA': 'Gln', 'CAG': 'Gln', 'GAA': 'Glu', 'GAG': 'Glu', 'GGT': 'Gly', 'GGC': 'Gly', 'GGA': 'Gly', 'GGG': 'Gly', 'CAT': 'His', 'CAC': 'His', 'ATT': 'Ile', 'ATC': 'Ile', 'ATA': 'Ile', 'CTT': 'Leu', 'CTC': 'Leu', 'CTA': 'Leu', 'CTG': 'Leu', 'TTA': 'Leu', 'TTG': 'Leu', 'AAA': 'Lys', 'AAG': 'Lys', 'ATG': 'Met', 'TTT': 'Phe', 'TTC': 'Phe', 'CCT': 'Pro', 'CCC': 'Pro', 'CCA': 'Pro', 'CCG': 'Pro', 'TCT': 'Ser', 'TCC': 'Ser', 'TCA': 'Ser', 'TCG': 'Ser', 'AGT': 'Ser', 'AGC': 'Ser', 'ACT': 'Thr', 'ACC': 'Thr', 'ACA': 'Thr', 'ACG': 'Thr', 'TGG': 'Trp', 'TAT': 'Tyr', 'TAC': 'Tyr', 'GTT': 'Val', 'GTC': 'Val', 'GTA': 'Val', 'GTG': 'Val', 'TAA': 'STOP', 'TGA': 'STOP', 'TAG': 'STOP' } export default codon2AA #+end_src ** amino acid to letter table :PROPERTIES: :VISIBILITY: folded :END: #+begin_src javascript :noweb yes :tangle aminoAcid2Code.mjs const aa2Code = { 'Ala': 'A', 'Arg': 'R', 'Asn': 'N', 'Asp': 'D', 'Cys': 'C', 'Gln': 'Q', 'Glu': 'E', 'Gly': 'G', 'His': 'H', 'Ile': 'I', 'Leu': 'L', 'Lys': 'K', 'Met': 'M', 'Phe': 'F', 'Pro': 'P', 'Ser': 'S', 'Thr': 'T', 'Trp': 'W', 'Tyr': 'Y', 'Val': 'V', 'STOP': '*' } export default aa2Code #+end_src ** logging #+begin_src javascript :noweb yes :tangle logging.mjs class Log { get elt() { if (!document.querySelector('#log')) { const elt = document.createElement('div'); elt.id = 'log'; document.body.appendChild(elt); } return document.querySelector('#log'); } clear() { while (this.elt.firstChild) { this.elt.removeChild(this.elt.firstChild); } } logAt(level, items) { const list = document.createElement('ul'); items.forEach((item) => { const msg = document.createElement('li'); msg.setAttribute('class', level); msg.innerText = item; list.appendChild(msg); }); this.elt.appendChild(list); } error(...items) { this.logAt('error', items); console.error.apply(null, items) } warn(...items) { this.logAt('warn', items); console.warn.apply(null, items) } info(...items) { console.info.apply(null, items) } debug(...items) { console.debug.apply(null, items) } }; Log.logger = new Log(); Log.clear = Log.logger.clear.bind(Log.logger); Log.error = Log.logger.error.bind(Log.logger); Log.warn = Log.logger.warn.bind(Log.logger); Log.info = Log.logger.info.bind(Log.logger); Log.debug = Log.logger.debug.bind(Log.logger); export default Log #+end_src