def main():
usage = 'usage: %prog [options] <model> <vcf_file>'
parser = OptionParser(usage)
parser.add_option('-c',
dest='slice_center',
default=None,
type='int',
help='Slice center positions [Default: %default]')
parser.add_option('-f',
dest='genome_fasta',
default='%s/data/hg38.fa' % os.environ['BASENJIDIR'],
help='Genome FASTA for sequences [Default: %default]')
parser.add_option('-n',
dest='norm_file',
default=None,
help='Normalize SAD scores')
parser.add_option(
'-o',
dest='out_dir',
default='sad',
help='Output directory for tables and plots [Default: %default]')
parser.add_option('-p',
dest='processes',
default=None,
type='int',
help='Number of processes, passed by multi script')
parser.add_option('--pseudo',
dest='log_pseudo',
default=1,
type='float',
help='Log2 pseudocount [Default: %default]')
parser.add_option(
'--rc',
dest='rc',
default=False,
action='store_true',
help=
'Average forward and reverse complement predictions [Default: %default]'
)
parser.add_option('--shifts',
dest='shifts',
default='0',
type='str',
help='Ensemble prediction shifts [Default: %default]')
parser.add_option('--species', dest='species', default='human')
parser.add_option(
'--stats',
dest='sad_stats',
default='SAD',
help='Comma-separated list of stats to save. [Default: %default]')
parser.add_option(
'-t',
dest='targets_file',
default=None,
type='str',
help='File specifying target indexes and labels in table format')
(options, args) = parser.parse_args()
if len(args) == 2:
# single worker
model_file = args[0]
vcf_file = args[1]
elif len(args) == 3:
# multi separate
options_pkl_file = args[0]
model_file = args[1]
vcf_file = args[2]
# save out dir
out_dir = options.out_dir
# load options
options_pkl = open(options_pkl_file, 'rb')
options = pickle.load(options_pkl)
options_pkl.close()
# update output directory
options.out_dir = out_dir
elif len(args) == 4:
# multi worker
options_pkl_file = args[0]
model_file = args[1]
vcf_file = args[2]
worker_index = int(args[3])
# load options
options_pkl = open(options_pkl_file, 'rb')
options = pickle.load(options_pkl)
options_pkl.close()
# update output directory
options.out_dir = '%s/job%d' % (options.out_dir, worker_index)
else:
parser.error('Must provide model and VCF file')
if not os.path.isdir(options.out_dir):
os.mkdir(options.out_dir)
options.shifts = [int(shift) for shift in options.shifts.split(',')]
options.sad_stats = options.sad_stats.split(',')
#################################################################
# read parameters and targets
if options.targets_file is None:
target_slice = None
else:
targets_df = pd.read_csv(options.targets_file, sep='\t', index_col=0)
target_ids = targets_df.identifier
target_labels = targets_df.description
target_slice = targets_df.index
#################################################################
# setup model
seqnn_model = tf.saved_model.load(model_file).model
# query num model targets
seq_length = seqnn_model.predict_on_batch.input_signature[0].shape[1]
null_1hot = np.zeros((1, seq_length, 4))
null_preds = seqnn_model.predict_on_batch(null_1hot)
null_preds = null_preds[options.species].numpy()
_, targets_length, num_targets = null_preds.shape
if options.targets_file is None:
target_ids = ['t%d' % ti for ti in range(num_targets)]
target_labels = [''] * len(target_ids)
#################################################################
# load SNPs
# filter for worker SNPs
if options.processes is not None:
# determine boundaries
num_snps = bvcf.vcf_count(vcf_file)
worker_bounds = np.linspace(0,
num_snps,
options.processes + 1,
dtype='int')
# read SNPs form VCF
snps = bvcf.vcf_snps(vcf_file,
start_i=worker_bounds[worker_index],
end_i=worker_bounds[worker_index + 1])
else:
# read SNPs form VCF
snps = bvcf.vcf_snps(vcf_file)
num_snps = len(snps)
# open genome FASTA
genome_open = pysam.Fastafile(options.genome_fasta)
# create SNP sequence generator
def snp_gen():
for snp in snps:
# get SNP sequences
snp_1hot_list = bvcf.snp_seq1(snp, seq_length, genome_open)
for snp_1hot in snp_1hot_list:
yield snp_1hot
#################################################################
# setup output
sad_out = initialize_output_h5(options.out_dir, options.sad_stats, snps,
target_ids, target_labels, targets_length)
#################################################################
# predict SNP scores, write output
# initialize predictions stream
preds_stream = PredStreamGen(seqnn_model,
snp_gen(),
rc=options.rc,
shifts=options.shifts,
slice_center=options.slice_center,
species=options.species)
# predictions index
pi = 0
for si in range(num_snps):
# get predictions
ref_preds = preds_stream[pi]
pi += 1
alt_preds = preds_stream[pi]
pi += 1
# process SNP
write_snp(ref_preds, alt_preds, sad_out, si, options.sad_stats,
options.log_pseudo)
# close genome
genome_open.close()
###################################################
# compute SAD distributions across variants
write_pct(sad_out, options.sad_stats)
sad_out.close()
def main():
usage = 'usage: %prog [options] <params_file> <model_file> <vcf_file>'
parser = OptionParser(usage)
parser.add_option(
'-c',
dest='center_pct',
default=0.25,
type='float',
help='Require clustered SNPs lie in center region [Default: %default]')
parser.add_option('-f',
dest='genome_fasta',
default='%s/data/hg19.fa' % os.environ['BASENJIDIR'],
help='Genome FASTA for sequences [Default: %default]')
parser.add_option(
'--flip',
dest='flip_ref',
default=False,
action='store_true',
help='Flip reference/alternate alleles when simple [Default: %default]'
)
parser.add_option('--local',
dest='local',
default=1024,
type='int',
help='Local SAD score [Default: %default]')
parser.add_option('-n',
dest='norm_file',
default=None,
help='Normalize SAD scores')
parser.add_option(
'-o',
dest='out_dir',
default='sad',
help='Output directory for tables and plots [Default: %default]')
parser.add_option('-p',
dest='processes',
default=None,
type='int',
help='Number of processes, passed by multi script')
parser.add_option('--pseudo',
dest='log_pseudo',
default=1,
type='float',
help='Log2 pseudocount [Default: %default]')
parser.add_option(
'--rc',
dest='rc',
default=False,
action='store_true',
help=
'Average forward and reverse complement predictions [Default: %default]'
)
parser.add_option('--shifts',
dest='shifts',
default='0',
type='str',
help='Ensemble prediction shifts [Default: %default]')
parser.add_option(
'--stats',
dest='sad_stats',
default='SAD',
help='Comma-separated list of stats to save. [Default: %default]')
parser.add_option(
'-t',
dest='targets_file',
default=None,
type='str',
help='File specifying target indexes and labels in table format')
parser.add_option(
'--ti',
dest='track_indexes',
default=None,
type='str',
help='Comma-separated list of target indexes to output BigWig tracks')
parser.add_option(
'--threads',
dest='threads',
default=False,
action='store_true',
help='Run CPU math and output in a separate thread [Default: %default]'
)
parser.add_option(
'-u',
dest='penultimate',
default=False,
action='store_true',
help='Compute SED in the penultimate layer [Default: %default]')
(options, args) = parser.parse_args()
if len(args) == 3:
# single worker
params_file = args[0]
model_file = args[1]
vcf_file = args[2]
elif len(args) == 5:
# multi worker
options_pkl_file = args[0]
params_file = args[1]
model_file = args[2]
vcf_file = args[3]
worker_index = int(args[4])
# load options
options_pkl = open(options_pkl_file, 'rb')
options = pickle.load(options_pkl)
options_pkl.close()
# update output directory
options.out_dir = '%s/job%d' % (options.out_dir, worker_index)
else:
parser.error(
'Must provide parameters and model files and QTL VCF file')
if not os.path.isdir(options.out_dir):
os.mkdir(options.out_dir)
if options.track_indexes is None:
options.track_indexes = []
else:
options.track_indexes = [
int(ti) for ti in options.track_indexes.split(',')
]
if not os.path.isdir('%s/tracks' % options.out_dir):
os.mkdir('%s/tracks' % options.out_dir)
options.shifts = [int(shift) for shift in options.shifts.split(',')]
options.sad_stats = options.sad_stats.split(',')
#################################################################
# read parameters and targets
# read model parameters
with open(params_file) as params_open:
params = json.load(params_open)
params_model = params['model']
params_train = params['train']
if options.targets_file is None:
target_slice = None
else:
targets_df = pd.read_csv(options.targets_file, sep='\t', index_col=0)
target_ids = targets_df.identifier
target_labels = targets_df.description
target_slice = targets_df.index
if options.penultimate:
parser.error('Not implemented for TF2')
#################################################################
# setup model
seqnn_model = seqnn.SeqNN(params_model)
seqnn_model.restore(model_file)
seqnn_model.build_slice(target_slice)
seqnn_model.build_ensemble(options.rc, options.shifts)
num_targets = seqnn_model.num_targets()
if options.targets_file is None:
target_ids = ['t%d' % ti for ti in range(num_targets)]
target_labels = [''] * len(target_ids)
#################################################################
# load SNPs
# filter for worker SNPs
if options.processes is not None:
# determine boundaries
num_snps = bvcf.vcf_count(vcf_file)
worker_bounds = np.linspace(0,
num_snps,
options.processes + 1,
dtype='int')
# read sorted SNPs from VCF
snps = bvcf.vcf_snps(vcf_file,
require_sorted=True,
flip_ref=options.flip_ref,
validate_ref_fasta=options.genome_fasta,
start_i=worker_bounds[worker_index],
end_i=worker_bounds[worker_index + 1])
else:
# read sorted SNPs from VCF
snps = bvcf.vcf_snps(vcf_file,
require_sorted=True,
flip_ref=options.flip_ref,
validate_ref_fasta=options.genome_fasta)
# cluster SNPs by position
snp_clusters = cluster_snps(snps, params_model['seq_length'],
options.center_pct)
# delimit sequence boundaries
[sc.delimit(params_model['seq_length']) for sc in snp_clusters]
# open genome FASTA
genome_open = pysam.Fastafile(options.genome_fasta)
# make SNP sequence generator
def snp_gen():
for sc in snp_clusters:
snp_1hot_list = sc.get_1hots(genome_open)
for snp_1hot in snp_1hot_list:
yield snp_1hot
#################################################################
# setup output
snp_flips = np.array([snp.flipped for snp in snps], dtype='bool')
sad_out = initialize_output_h5(options.out_dir, options.sad_stats, snps,
target_ids, target_labels)
if options.threads:
snp_threads = []
snp_queue = Queue()
for i in range(1):
sw = SNPWorker(snp_queue, sad_out, options.sad_stats,
options.log_pseudo)
sw.start()
snp_threads.append(sw)
#################################################################
# predict SNP scores, write output
# initialize predictions stream
preds_stream = stream.PredStreamGen(seqnn_model, snp_gen(),
params['train']['batch_size'])
# predictions index
pi = 0
# SNP index
si = 0
for snp_cluster in snp_clusters:
ref_preds = preds_stream[pi]
pi += 1
for snp in snp_cluster.snps:
# print(snp, flush=True)
alt_preds = preds_stream[pi]
pi += 1
if snp_flips[si]:
ref_preds, alt_preds = alt_preds, ref_preds
if options.threads:
# queue SNP
snp_queue.put((ref_preds, alt_preds, si))
else:
# process SNP
write_snp(ref_preds, alt_preds, sad_out, si, options.sad_stats,
options.log_pseudo)
# update SNP index
si += 1
# finish queue
if options.threads:
print('Waiting for threads to finish.', flush=True)
snp_queue.join()
# close genome
genome_open.close()
###################################################
# compute SAD distributions across variants
write_pct(sad_out, options.sad_stats)
sad_out.close()