# Calling Cards Region Quantification # Quantifies enrichment of insertions/hops in genomic regions # # This script: # 1. Counts insertions overlapping each genomic region (experiment and background) # 2. Calculates enrichment scores # 3. Computes Poisson and hypergeometric p-values # # Works with any data in BED3+ format (chr, start, end, ...) # For calling cards: each insertion is counted once regardless of depth # # COORDINATE SYSTEMS: # - Input BED files are assumed to be 0-indexed, half-open [start, end) # - GenomicRanges uses 1-indexed, closed [start, end] # - Conversion: GR_start = BED_start + 1, GR_end = BED_end # # Usage: # Rscript quantify_regions.R library(tidyverse) library(GenomicRanges) # Statistical Functions --------------------------------------------------- #' Calculate enrichment (calling cards effect) #' #' @param total_background_hops Total number of hops in background (scalar or vector) #' @param total_experiment_hops Total number of hops in experiment (scalar or vector) #' @param background_hops Number of hops in background per region (vector) #' @param experiment_hops Number of hops in experiment per region (vector) #' @param pseudocount Pseudocount to avoid division by zero (default: 0.1) #' @return Enrichment values calculate_enrichment <- function(total_background_hops, total_experiment_hops, background_hops, experiment_hops, pseudocount = 0.1) { # Input validation if (!all(is.numeric(c( total_background_hops, total_experiment_hops, background_hops, experiment_hops )))) { stop("All inputs must be numeric") } # Get the length of the region vectors n_regions <- length(background_hops) # Ensure experiment_hops is same length as background_hops if (length(experiment_hops) != n_regions) { stop("background_hops and experiment_hops must be the same length") } # Recycle scalar totals to match region length if needed if (length(total_background_hops) == 1) { total_background_hops <- rep(total_background_hops, n_regions) } if (length(total_experiment_hops) == 1) { total_experiment_hops <- rep(total_experiment_hops, n_regions) } # Now check all are same length if (length(total_background_hops) != n_regions || length(total_experiment_hops) != n_regions) { stop("All input vectors must be the same length or scalars") } # Calculate enrichment numerator <- experiment_hops / total_experiment_hops denominator <- (background_hops + pseudocount) / total_background_hops enrichment <- numerator / denominator # Check for invalid values if (any(enrichment < 0, na.rm = TRUE)) { stop("Enrichment values must be non-negative") } if (any(is.na(enrichment))) { stop("Enrichment values must not be NA") } if (any(is.infinite(enrichment))) { stop("Enrichment values must not be infinite") } return(enrichment) } #' Calculate Poisson p-values #' #' @param total_background_hops Total number of hops in background (scalar or vector) #' @param total_experiment_hops Total number of hops in experiment (scalar or vector) #' @param background_hops Number of hops in background per region (vector) #' @param experiment_hops Number of hops in experiment per region (vector) #' @param pseudocount Pseudocount for lambda calculation (default: 0.1) #' @param ... additional arguments to `ppois`. note that lower tail is set to FALSE #' already #' @return Poisson p-values calculate_poisson_pval <- function(total_background_hops, total_experiment_hops, background_hops, experiment_hops, pseudocount = 0.1, ...) { # Input validation if (!all(is.numeric(c( total_background_hops, total_experiment_hops, background_hops, experiment_hops )))) { stop("All inputs must be numeric") } # Get the length of the region vectors n_regions <- length(background_hops) # Ensure experiment_hops is same length as background_hops if (length(experiment_hops) != n_regions) { stop("background_hops and experiment_hops must be the same length") } # Recycle scalar totals to match region length if needed if (length(total_background_hops) == 1) { total_background_hops <- rep(total_background_hops, n_regions) } if (length(total_experiment_hops) == 1) { total_experiment_hops <- rep(total_experiment_hops, n_regions) } # Now check all are same length if (length(total_background_hops) != n_regions || length(total_experiment_hops) != n_regions) { stop("All input vectors must be the same length or scalars") } # Calculate hop ratio hop_ratio <- total_experiment_hops / total_background_hops # Calculate expected number of hops (mu/lambda parameter) # Add pseudocount to avoid mu = 0 mu <- (background_hops + pseudocount) * hop_ratio # Observed hops in experiment x <- experiment_hops # Calculate p-value: P(X >= x) = 1 - P(X < x) = 1 - P(X <= x-1) # This is equivalent to: 1 - CDF(x) + PMF(x) # Using the upper tail directly with lower.tail = FALSE pval <- ppois(x - 1, lambda = mu, lower.tail = FALSE, ...) return(pval) } #' Calculate hypergeometric p-values #' #' @param total_background_hops Total number of hops in background (scalar or vector) #' @param total_experiment_hops Total number of hops in experiment (scalar or vector) #' @param background_hops Number of hops in background per region (vector) #' @param experiment_hops Number of hops in experiment per region (vector) #' @param ... additional arguments to phyper. Note that lower tail is set to #' false already #' @return Hypergeometric p-values calculate_hypergeom_pval <- function(total_background_hops, total_experiment_hops, background_hops, experiment_hops, ...) { # Input validation if (!all(is.numeric(c( total_background_hops, total_experiment_hops, background_hops, experiment_hops )))) { stop("All inputs must be numeric") } # Get the length of the region vectors n_regions <- length(background_hops) # Ensure experiment_hops is same length as background_hops if (length(experiment_hops) != n_regions) { stop("background_hops and experiment_hops must be the same length") } # Recycle scalar totals to match region length if needed if (length(total_background_hops) == 1) { total_background_hops <- rep(total_background_hops, n_regions) } if (length(total_experiment_hops) == 1) { total_experiment_hops <- rep(total_experiment_hops, n_regions) } # Now check all are same length if (length(total_background_hops) != n_regions || length(total_experiment_hops) != n_regions) { stop("All input vectors must be the same length or scalars") } # Hypergeometric parameters # M: total number of balls (total hops) M <- total_background_hops + total_experiment_hops # n: number of white balls (experiment hops) n <- total_experiment_hops # N: number of draws (hops in region) N <- background_hops + experiment_hops # x: number of white balls drawn (experiment hops in region) - 1 for upper tail x <- experiment_hops - 1 # Handle edge cases valid <- (M >= 1) & (N >= 1) pval <- rep(1, length(M)) # Calculate p-value for valid cases: P(X >= x) = 1 - P(X <= x-1) if (any(valid)) { pval[valid] <- phyper(x[valid], n[valid], M[valid] - n[valid], N[valid], lower.tail = FALSE, ... ) } return(pval) } # GRanges Conversion Functions -------------------------------------------- #' Convert BED format data frame to GRanges #' #' Handles coordinate system conversion from 0-indexed half-open BED format #' to 1-indexed closed GenomicRanges format #' #' @param bed_df Data frame with chr, start, end columns in BED format (0-indexed, half-open) #' @param zero_indexed Logical, whether input is 0-indexed (default: TRUE) #' @return GRanges object bed_to_granges <- function(bed_df, zero_indexed = TRUE) { if (!all(c("chr", "start", "end") %in% names(bed_df))) { stop("bed_df must have columns: chr, start, end") } # Convert from 0-indexed half-open [start, end) to 1-indexed closed [start, end] if (zero_indexed) { gr_start <- bed_df$start + 1 gr_end <- bed_df$end } else { gr_start <- bed_df$start gr_end <- bed_df$end } # Create GRanges object (strand-agnostic for calling cards) gr <- GRanges( seqnames = bed_df$chr, ranges = IRanges(start = gr_start, end = gr_end), strand = "*" ) # Add any additional metadata columns extra_cols <- setdiff(names(bed_df), c("chr", "start", "end", "strand")) if (length(extra_cols) > 0) { mcols(gr) <- bed_df[, extra_cols, drop = FALSE] } return(gr) } #' Deduplicate insertions in GRanges object #' #' For calling cards, if an insertion is found at the same coordinate, #' only one record is retained #' #' @param gr GRanges object #' @return Deduplicated GRanges object deduplicate_granges <- function(gr) { # Find unique ranges (ignores strand and metadata) unique_ranges <- !duplicated(granges(gr)) gr[unique_ranges] } #' Count overlaps between insertions and regions #' #' @param insertions_gr GRanges object with insertions #' @param regions_gr GRanges object with regions #' @param deduplicate Whether to deduplicate insertions (default: TRUE) #' @return Integer vector of overlap counts per region count_overlaps <- function(insertions_gr, regions_gr, deduplicate = TRUE) { # Deduplicate if requested if (deduplicate) { n_before <- length(insertions_gr) insertions_gr <- deduplicate_granges(insertions_gr) n_after <- length(insertions_gr) if (n_before != n_after) { message( " Deduplicated: ", n_before, " -> ", n_after, " (removed ", n_before - n_after, " duplicates)" ) } } # Count overlaps per region # countOverlaps returns an integer vector with one element per region counts <- countOverlaps(regions_gr, insertions_gr) return(counts) } # Main Analysis Function -------------------------------------------------- #' Call peaks/quantify regions using calling cards approach #' #' @param experiment_gr GRanges object with experiment insertions #' @param background_gr GRanges object with background insertions #' @param regions_gr GRanges object with regions to quantify #' @param deduplicate_experiment Whether to deduplicate experiment insertions (default: TRUE) #' @param pseudocount Pseudocount for calculations (default: 0.1) #' @return GRanges object with regions and statistics as metadata columns enrichment_analysis <- function(experiment_gr, background_gr, regions_gr, deduplicate_experiment = TRUE, pseudocount = 0.1) { message("Starting enrichment analysis...") # Validate inputs if (!inherits(experiment_gr, "GRanges")) { stop("experiment_gr must be a GRanges object") } if (!inherits(background_gr, "GRanges")) { stop("background_gr must be a GRanges object") } if (!inherits(regions_gr, "GRanges")) { stop("regions_gr must be a GRanges object") } # Count overlaps for experiment (with deduplication if requested) message("Counting experiment overlaps...") if (deduplicate_experiment) { message(" Deduplication: ON") } else { message(" Deduplication: OFF") } experiment_counts <- count_overlaps( experiment_gr, regions_gr, deduplicate = deduplicate_experiment ) # Count overlaps for background (never deduplicated) message("Counting background overlaps...") message(" Deduplication: OFF (background should not be deduplicated)") background_counts <- count_overlaps( background_gr, regions_gr, deduplicate = FALSE ) # Calculate total hops AFTER any deduplication if (deduplicate_experiment) { experiment_gr_dedup <- deduplicate_granges(experiment_gr) total_experiment_hops <- length(experiment_gr_dedup) } else { total_experiment_hops <- length(experiment_gr) } total_background_hops <- length(background_gr) message("Total experiment hops: ", total_experiment_hops) message("Total background hops: ", total_background_hops) if (total_experiment_hops == 0) { stop("Experiment data is empty") } if (total_background_hops == 0) { stop("Background data is empty") } # Add counts and totals as metadata columns mcols(regions_gr)$experiment_hops <- as.integer(experiment_counts) mcols(regions_gr)$background_hops <- as.integer(background_counts) mcols(regions_gr)$total_experiment_hops <- as.integer(total_experiment_hops) mcols(regions_gr)$total_background_hops <- as.integer(total_background_hops) # Calculate statistics message("Calculating enrichment scores...") mcols(regions_gr)$callingcards_enrichment <- calculate_enrichment( total_background_hops = total_background_hops, total_experiment_hops = total_experiment_hops, background_hops = background_counts, experiment_hops = experiment_counts, pseudocount = pseudocount ) message("Calculating Poisson p-values...") mcols(regions_gr)$poisson_pval <- calculate_poisson_pval( total_background_hops = total_background_hops, total_experiment_hops = total_experiment_hops, background_hops = background_counts, experiment_hops = experiment_counts, pseudocount = pseudocount ) message("Calculating log Poisson p-values...") mcols(regions_gr)$log_poisson_pval <- calculate_poisson_pval( total_background_hops = total_background_hops, total_experiment_hops = total_experiment_hops, background_hops = background_counts, experiment_hops = experiment_counts, pseudocount = pseudocount, log.p = TRUE ) message("Calculating hypergeometric p-values...") mcols(regions_gr)$hypergeometric_pval <- calculate_hypergeom_pval( total_background_hops = total_background_hops, total_experiment_hops = total_experiment_hops, background_hops = background_counts, experiment_hops = experiment_counts ) message("Calculating log hypergeometric p-values...") mcols(regions_gr)$log_hypergeometric_pval <- calculate_hypergeom_pval( total_background_hops = total_background_hops, total_experiment_hops = total_experiment_hops, background_hops = background_counts, experiment_hops = experiment_counts, log.p = TRUE ) # Calculate adjusted p-values message("Calculating adjusted p-values...") mcols(regions_gr)$poisson_qval <- p.adjust(mcols(regions_gr)$poisson_pval, method = "fdr") mcols(regions_gr)$hypergeometric_qval <- p.adjust(mcols(regions_gr)$hypergeometric_pval, method = "fdr") message("Analysis complete!") return(regions_gr) } # Example Usage ----------------------------------------------------------- PROMOTERS <- "intergenic" genomic_features <- arrow::read_parquet("~/projects/huggingface/yeast_genome_resources/brentlab_features.parquet") genome_map_replicate_ds <- arrow::open_dataset("~/projects/huggingface/callingcards/genome_map", unify_schemas = TRUE) genome_map_replicate_meta <- arrow::read_parquet("~/projects/huggingface/callingcards/genome_map_meta.parquet") background_gr <- read_tsv("~/projects/huggingface/callingcards/adh1_background_ucsc.qbed") |> mutate( id = "adh1_bg", score = scales::rescale(depth, to = c(1, 1000)) ) |> dplyr::select(chr, start, end, id, score, strand) |> bed_to_granges() regions_list <- list( yiming = read_tsv("~/projects/huggingface/yeast_genome_resources/yiming_promoters.bed", col_names = c("chr", "start", "end", "locus_tag", "score", "strand") ) %>% bed_to_granges(), mindel = arrow::read_parquet("~/projects/huggingface/yeast_genome_resources/mindel_promoters.parquet", col_names = c("chr", "start", "end", "locus_tag", "score", "strand") ) %>% bed_to_granges(), start_codon_500bp = rtracklayer::import("~/projects/huggingface/yeast_genome_resources/start_codon_500bp_upstream_promoters.bed"), intergenic = rtracklayer::import("~/projects/huggingface/yeast_genome_resources/intergenic_regions_5_1.bed") ) # rename name to target_locus_tag colnames(GenomicRanges::mcols(regions_list$yiming))[1] <- "target_locus_tag" colnames(GenomicRanges::mcols(regions_list$start_codon_500bp))[1] <- "target_locus_tag" intergenic_meta <- read_csv("~/projects/huggingface/yeast_genome_resources/intergenic_regions_metadata_5_1.csv") |> dplyr::select(ir_name, chr, start, end, feature_left, feature_right) |> pivot_longer(-c(ir_name, chr, start, end), names_to = "side", values_to = "target_locus_tag") |> left_join(dplyr::select(genomic_features, target_locus_tag = locus_tag, target_symbol = symbol, target_strand = strand, target_start = start, target_end = end )) |> filter(chr != "chrM") |> filter(!is.na(target_strand)) |> mutate(valid = (target_strand == "-" & target_end <= start) | (target_strand == "+" & target_start >= end)) |> filter(valid) regions_gr <- regions_list[[PROMOTERS]] # removes AAD6 and AAD16 from mindel. pseudogene no longer in annotations if (PROMOTERS != "intergenic") { # really just for mindel regions_gr <- regions_gr[!is.na(regions_gr$target_locus_tag)] } else if (PROMOTERS == "intergenic") { regions_gr <- regions_gr[regions_gr$name %in% unique(intergenic_meta$ir_name)] } library(parallel) n_cores <- 10 results_replicates <- mclapply( genome_map_replicate_meta$id, \(x) enrichment_analysis( experiment_gr = genome_map_replicate_ds |> filter(id == x) |> collect() |> dplyr::rename(score = depth) |> relocate(chr, start, end, id, score, strand) |> mutate(depth = scales::rescale(score, to = c(1, 1000))) |> bed_to_granges(), background_gr = background_gr, regions_gr = regions_gr, deduplicate_experiment = TRUE, pseudocount = 0.1 ), mc.cores = n_cores ) names(results_replicates) <- genome_map_replicate_meta$id if (PROMOTERS == "intergenic") { results_replicates_df <- bind_rows(map(results_replicates, as_tibble), .id = "genome_map_id") |> mutate(genome_map_id = as.integer(genome_map_id)) |> dplyr::rename(ir_name = name) |> left_join(dplyr::select(intergenic_meta, ir_name, target_locus_tag, target_symbol), relationship = "many-to-many" ) |> left_join(genome_map_replicate_meta, by = c("genome_map_id" = "id") ) |> select( genome_map_id, batch, target_locus_tag, target_symbol, ir_name, experiment_hops, background_hops, total_background_hops, total_experiment_hops, callingcards_enrichment, poisson_pval, log_poisson_pval, poisson_qval, hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval ) |> mutate(genome_map_id = as.integer(genome_map_id)) # arrow::write_dataset( # results_replicates_df, # path = "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_intergenic", # format = "parquet", # partitioning = c("batch"), # existing_data_behavior = "overwrite", # compression = "zstd", # write_statistics = TRUE, # use_dictionary = c( # genome_map_id = TRUE # ) # ) } else if (PROMOTERS == "start_codon_500bp") { results_replicates_df <- bind_rows(map(results_replicates, as_tibble), .id = "genome_map_id") |> mutate(genome_map_id = as.integer(genome_map_id)) |> left_join(dplyr::select(genomic_features, target_locus_tag = locus_tag, target_symbol = symbol)) |> left_join(genome_map_replicate_meta, by = c("genome_map_id" = "id") ) |> select( genome_map_id, batch, target_locus_tag, target_symbol, experiment_hops, background_hops, total_background_hops, total_experiment_hops, callingcards_enrichment, poisson_pval, log_poisson_pval, poisson_qval, hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval ) |> mutate(genome_map_id = as.integer(genome_map_id)) # arrow::write_dataset( # results_replicates_df, # path = "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_start_codon_500", # format = "parquet", # partitioning = c("batch"), # existing_data_behavior = "overwrite", # compression = "zstd", # write_statistics = TRUE, # use_dictionary = c( # genome_map_id = TRUE # ) # ) } # creating the combined data for a new set # NOTE: THERE IS AN ERROR HERE. FOR THE SINGLETONS, THE ID IS THE # ANNOTATED_FEATURE ID, NOT THE GENOME_MAP_ID # as_2026 <- arrow::read_parquet("~/projects/huggingface/callingcards/deprecated/2026_analysis_set.parquet") # annotated_feature_meta <- arrow::read_parquet("~/projects/huggingface/callingcards/annotated_feature_meta.parquet") # as_2026_singles <- as_2026 |> # filter(str_detect(gm_id, "-", negate = TRUE)) |> # dplyr::rename(id = gm_id) |> # mutate(id = as.numeric(id)) |> # left_join( # annotated_feature_meta |> # dplyr::select(id, gm_id = genome_map_id) # ) |> # dplyr::relocate(id, gm_id) |> # dplyr::select(-id) |> # mutate(gm_id = as.character(gm_id)) # # as_2026_revised <- as_2026 |> # filter(str_detect(gm_id, "-")) |> # bind_rows(as_2026_singles) # # arrow::write_parquet(as_2026_revised, "~/projects/huggingface/callingcards/2026_analysis_set.parquet") as_2026 <- arrow::read_parquet("~/projects/huggingface/callingcards/2026_analysis_set.parquet") passing_rep_gm_ids <- unlist(map(unique(as_2026$gm_id), str_split, "-", simplify = TRUE)) genome_map_replicate_meta_combined <- genome_map_replicate_meta |> filter(id %in% passing_rep_gm_ids) |> group_by(regulator_locus_tag, regulator_symbol, condition) |> mutate(id = as.character(id)) |> reframe(combined_id = if (n() > 1) paste(id, collapse = "-") else id) # # Run analysis with deduplication (default for calling cards) results_combined <- map(genome_map_replicate_meta_combined$combined_id, ~ { # get list of ids (might just be 1) ids <- as.integer(strsplit(.x, "-")[[1]]) ids <- ids[!is.na(ids)] # for each id, produce a GRanges combined_passing_set <- map(ids, ~ { genome_map_replicate_ds |> filter(id == .x) |> collect() |> dplyr::rename(score = depth) |> relocate(chr, start, end, id, score, strand) |> mutate(depth = scales::rescale(score, to = c(1, 1000))) |> bed_to_granges() |> deduplicate_granges() }) # combine the list into a single gr combined_gr <- do.call(c, combined_passing_set) enrichment_analysis( experiment_gr = combined_gr, background_gr = background_gr, regions_gr = regions_gr, deduplicate_experiment = FALSE, pseudocount = 0.1 ) }) names(results_combined) <- genome_map_replicate_meta_combined$combined_id if (PROMOTERS == "intergenic") { results_combined_analysis_df <- bind_rows(map(results_combined, as_tibble), .id = "combined_id") |> dplyr::rename(ir_name = name) |> left_join(dplyr::select(intergenic_meta, ir_name, target_locus_tag, target_symbol), relationship = "many-to-many" ) |> left_join(genome_map_replicate_meta_combined) |> select( gm_id = combined_id, regulator_locus_tag, regulator_symbol, condition, target_locus_tag, target_symbol, experiment_hops, background_hops, total_background_hops, total_experiment_hops, callingcards_enrichment, poisson_pval, log_poisson_pval, poisson_qval, hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval ) |> filter(condition == "standard") results_combined_analysis_df |> ungroup() |> dplyr::select(-condition) |> arrange(regulator_locus_tag) |> arrow::write_parquet( "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_intergenic_analysis.parquet", compression = "zstd", write_statistics = TRUE, use_dictionary = TRUE ) } else if (PROMOTERS == "start_codon_500bp") { results_combined_analysis_df <- bind_rows(map(results_combined, as_tibble), .id = "combined_id") |> left_join(dplyr::select(genomic_features, target_locus_tag = locus_tag, target_symbol = symbol)) |> left_join(genome_map_replicate_meta_combined) |> select( gm_id = combined_id, regulator_locus_tag, regulator_symbol, condition, target_locus_tag, target_symbol, experiment_hops, background_hops, total_background_hops, total_experiment_hops, callingcards_enrichment, poisson_pval, log_poisson_pval, poisson_qval, hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval ) |> filter(condition == "standard") results_combined_analysis_df |> ungroup() |> dplyr::select(-condition) |> arrange(regulator_locus_tag) |> arrow::write_parquet( "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_start_codon_500bp_analysis.parquet", compression = "zstd", write_statistics = TRUE, use_dictionary = TRUE ) } else if (PROMOTERS == "yiming") { results_combined_analysis_df <- bind_rows(map(results_combined, as_tibble), .id = "combined_id") |> left_join(dplyr::select(genomic_features, target_locus_tag = locus_tag, target_symbol = symbol)) |> left_join(genome_map_replicate_meta_combined) |> select( gm_id = combined_id, regulator_locus_tag, regulator_symbol, condition, target_locus_tag, target_symbol, experiment_hops, background_hops, total_background_hops, total_experiment_hops, callingcards_enrichment, poisson_pval, log_poisson_pval, poisson_qval, hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval ) |> filter(condition == "standard") out <- results_combined_analysis_df |> ungroup() |> dplyr::select(-condition) |> arrange(regulator_locus_tag) out |> arrow::write_parquet( "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_yiming_analysis.parquet", compression = "zstd", write_statistics = TRUE, use_dictionary = TRUE ) } else if (PROMOTERS == "mindel") { results_combined_analysis_df <- bind_rows(map(results_combined, as_tibble), .id = "combined_id") |> left_join(dplyr::select(genomic_features, target_locus_tag = locus_tag, target_symbol = symbol)) |> left_join(genome_map_replicate_meta_combined) |> select( gm_id = combined_id, regulator_locus_tag, regulator_symbol, condition, target_locus_tag, target_symbol, experiment_hops, background_hops, total_background_hops, total_experiment_hops, callingcards_enrichment, poisson_pval, log_poisson_pval, poisson_qval, hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval ) |> filter(condition == "standard") out <- results_combined_analysis_df |> ungroup() |> dplyr::select(-condition) |> arrange(regulator_locus_tag) out |> arrow::write_parquet( "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_mindel_analysis.parquet", compression = "zstd", write_statistics = TRUE, use_dictionary = TRUE ) }