MiXCR v4.6.0

🖇️ Combined Heavy+Light Somatic Hypermutation Trees from Single-Cell data

• A special step is added in findShmTrees to combine heavy and light SHM trees utilizing information added to clonotypes by groupClones command. Nodes in resulting tree will contain both light and heavy chains. If there is no connection to a clone from a companion chain, a reconstructed sequence will be added.
• Behaviour can be disabled with --dont-combine-tree-by-cells option to reconstruct separate heavy and light SHM trees
• Added exportShmSingleCellTrees command that export one node per line. It there is several roots in a tree, data will be exported in a different columns.
• Added -subtreeId to tree exports to differentiate part of trees from different chains
• exportShmTreesWithNodes and exportShmTrees commands will export subtrees with different chains at separate rows.

🚀 Other major upgrades

Changes in groupClones command

• Previous algorithm replaced with a new one that have better way of working with contamination, can detect multi-mappers (when one cell barcode marks two different cells) and can work with non-functional clones
• Some clones are now explicitly marked as contamination. This information is available as a separate label in exportClones in groupId column. Such clones can be filtered out from export by --filter-out-group-types contamination
• More important algorithm performance metrics are added to the report
• Fix for behaviour leading to clones with undefiened group being split by cell barcodes

New characteristics in SHM trees exports

• -subtreeId for determination of different chains in the same tree
• -numberOfClonesInTree [forChain] Number of uniq clones in the SHM tree.
• -numberOfNodesWithClones Number of nodes with clones, i.e. nodes with different clone sequences.
• -totalReadsCountInTree [forChain] Total sum of read counts of clones in the SHM tree.
• -totalUniqueTagCountInTree (Molecule|Cell|Sample) [forChain] Total count of unique tags in the SHM tree with specified type.
• -chains Chain type of the tree
• -treeHeight Height of the tree
• -vGene, -jGene, -vFamily, -jFamily - in previous version thous were exported only for nodes with clones
• -vBestIdentityPercent, -jBestIdentityPercent, -isOOF and -isProductive now exported for reconstructed nodes too

New characteristics in clonotype export

• -aaLength and -allAALength is available alongside -nLength and -allNLength
• -aaMutationsRate is available alongside -nMutationsRate
• Added optional arg germline in -nFeature, -aaFeature, -nLength, -aaLength in exportClones, exportAlignments and exportCloneGroups. It allows to export a sequence of the germline instead of a sequence of the gene.
• For all mutation exports (excluding -mutationsDetailed ) added optional filter by mutation type: ... [(substitutions|indels|inserts|deletions)]
• Added -nMutationsCount, -aaMutationsCount, -allNMutationsCount, -allAAMutationsCount for all relatable exports
• For mutation exports in exportShmTreesWithNodes (germline|mrca|parent) option is now optional. Will be export mutations from germline by default
• Added --export-clone-groups-sort-chains-by mixin
• Nucleotide mutations now could be exported for features that contain VCDR3Part, DCDR3Part or JCDR3Part
• Now -nLength, -nMutationsCount, -nMutationsRate can be calculated for multiple gene features (e.g. -nMutationsRate VRegionTrimmed,JRegionTrimmed)
• Added --export-clone-groups-sort-chains-by mixin with type of sorting of clones for determination of the primary and the secondary chains. It applies to exportCloneGroups command. By default, it's Auto (by UMI if it's available, by Read otherwise; previous default value was Read)
• Added --filter-out-group-types mixin to filter-out clones having certain clone group assignment kind: found, undefined or contamination. It applies to exportClones command
• Now exportCloneGroups by default will export groups in separate files for IG, TRAB, TRGD and mixed. This behaviour could be switched off by using --reset-export-clone-table-splitting or single --export-clone-groups-for-cell-type. In case of several --export-clone-groups-for-cell-type every cell type will be exported in separate file.
• In case of --export-clone-groups-for-cell-type in exportCloneGroups all mixed or unmatched groups will be filtered out.
• Added read and Molecule fraction columns to single cell exportClones output.

🧬 Reference library upgrades

• Previous TRAD meta-chain split into TRA and TRD as it should be. Chain assignment for clonotypes based on J genes.
• Rebuild allelic reference for human IGH, TRB, TRA and TRD chains. Now allelic names correspond to the IUIS nomenclature.
• Human IGK Vend coordinates corrected.
• UTR5Begin coordinates added to the following mouse genes: IGKV23-1, IGKV20-101-2, IGKV14-130, IGKV8-28, TRGV2

📚 Preset updates

• The milab-human-rna-tcr-umi-race preset has been updated: now clones are assembled by default based on the CDR3, in line with the manufacturer's recommended read length.
• The flairr-seq-bcr preset has been updated: now the preset sets species to human by default according to a built-in tag pattern with primer sequences.
• The following presets have been added to cover Ivivoscribe assay panels: invivoscribe-human-dna-trg-lymphotrack,invivoscribe-human-dna-trb-lymphotrack, invivoscribe-human-dna-igk-lymphotrack,invivoscribe-human-dna-ighv-leader-lymphotrack,invivoscribe-human-dna-igh-fr3-lymphotrack, invivoscribe-human-dna-igh-fr2-lymphotrack,invivoscribe-human-dna-igh-fr1-lymphotrack,invivoscribe-human-dna-igh-fr123-lymphotrack.
• The following presets have been added for mouse Thermofisher assays: thermofisher-mouse-rna-tcb-ampliseq-sr,thermofisher-mouse-dna-tcb-ampliseq-sr,thermofisher-mouse-rna-igh-ampliseq-sr,thermofisher-mouse-dna-igh-ampliseq-sr.
• Preset for SMARTer Human scTCR a/b Profiling Kit: takara-sc-human-rna-tcr-smarter
• The milab-human-rna-ig-umi-multiplex preset has been updated: the pattern now trims fewer nucleotides, which facilitates CDR1 identification. The splits by V and J genes have been removed as redundant due to the full-length assembling feature.

🛠️ Minor improvements & fixes

• More strict Combining trees step in findShmTrees command
• Better calculation of indel mutations between nodes in process of building shm trees
• Increased percent of successful alignment-aided overlaps by removing unnecessary overlap region quality sum threshold
• Impossible export of germline sequence for VJJunction in shmTrees exports now produces an error
• Parameter validation fix in -nMutationsRate
• Fix for -nMutationsRate if region is not covered for the clone
• Fix for the formal of exportAlignmentsPretty broken in the previous version
• Fix for IllegalArgumentException in exportAlignmentsPretty for cases where translation can't be performed
• Fix error for analyze executed with -f and --output-not-used-reads at the same time
• Resolutions of wildcards are excluded from calculation of -nMutationsRate for CDR3 in exportShmTreesWithNodes
• Fix OutOfMemory exception in command extend with .vdjca input
• In findShmTrees filter for productive only clones now check for stop codons in all features, not only in CDR3
• Change default value for filter for productive clones in findShmTrees to false (was true before)
• Add option --productive-only to findShmTrees
• Fixed parsing of --export-clone-groups-for-cell-type parameter
• Fixed usage of slice command on clnx files that weren't ordered by id.
• In slice now default behaviour is to keep original ids. Previous behaviour available with --reassign-ids option
• Fixed parsing of composite gene features with offsets like --assemble-clonotypes-by [VDJRegion,CBegin(0,10)]
• Fixed parent directory creation for output of exportClonesOverlap
• Fixed exportAirr in case of a clone with CDR3 that don't have VCDR3Part and JCDR3Part
• Optimize calculation of ranks in clone set. Speeds up export with tags and several other places.
• Added clone_id column in exportAirr
• Fixed exportClones in case of splitting file by tag:... if there is a clone that have several tags of requested level
• Fixed calculation of -nMutationsCount, -nMutationsRate, -aaMutationsCount and -aaMutationsRate. Previously in some cases it was calculated on different region, from what was requested.
• Added CellBarcodesWithFoundGroups for groupClones QC checks
• New filter --no-feature in exportAlignmentsPretty
• Fixed reporting in align, now coverage takes into account alignment-aided overlap

❗ Breaking changes

• Option --build-from <path> was removed from findShmTrees command

Deprecations of export options

• -lengthOf now is deprecated, use -nLength instead
• -allLengthOf now is deprecated, use -allNLength instead
• -mutationRate now is deprecated, use -nMutationsRate instead