int splicing_simulate_reads(const splicing_gff_t *gff, int gene,
const splicing_vector_t *expression,
int noreads, int readLength,
splicing_vector_int_t *isoform,
splicing_vector_int_t *position,
splicing_strvector_t *cigar,
splicing_vector_t *sample_prob) {
size_t i, p, noiso, goodiso=0, nogenes;
splicing_vector_int_t effisolen;
splicing_vector_t sampleprob;
double rand, sumpsi=0.0;
splicing_vector_int_t exstart, exend, exidx;
SPLICING_CHECK(splicing_gff_nogenes(gff, &nogenes));
if (gene < 0 || gene >= nogenes) {
SPLICING_ERROR("Invalid gene id", SPLICING_EINVAL);
}
/* TODO: more error checks */
SPLICING_CHECK(splicing_gff_noiso_one(gff, gene, &noiso));
SPLICING_CHECK(splicing_vector_int_init(&effisolen, noiso));
SPLICING_FINALLY(splicing_vector_int_destroy, &effisolen);
SPLICING_CHECK(splicing_vector_init(&sampleprob, noiso));
SPLICING_FINALLY(splicing_vector_destroy, &sampleprob);
SPLICING_CHECK(splicing_vector_int_resize(isoform, noreads));
SPLICING_CHECK(splicing_gff_isolength_one(gff, gene, &effisolen));
for (i=0; i<noiso; i++) {
int l=VECTOR(effisolen)[i]-readLength+1;
VECTOR(effisolen)[i] = l > 0 ? l : 0;
VECTOR(sampleprob)[i] = VECTOR(*expression)[i] * VECTOR(effisolen)[i];
if (VECTOR(sampleprob)[i] != 0) { goodiso++; }
sumpsi += VECTOR(sampleprob)[i];
}
if (goodiso==0) {
SPLICING_ERROR("No isoform is possible", SPLICING_FAILURE);
}
if (sample_prob) {
SPLICING_CHECK(splicing_vector_update(sample_prob, &sampleprob));
}
for (i=1; i<noiso; i++) {
VECTOR(sampleprob)[i] += VECTOR(sampleprob)[i-1];
}
for (i=0; i<noreads; i++) {
int w;
if (noiso==1) {
w=0;
} else if (noiso==2) {
rand = RNG_UNIF01() * sumpsi;
w = (rand < VECTOR(sampleprob)[0]) ? 0 : 1;
} else {
rand = RNG_UNIF01() * sumpsi;
for (w=0; rand > VECTOR(sampleprob)[w]; w++) ;
}
VECTOR(*isoform)[i]=w;
}
splicing_vector_destroy(&sampleprob);
SPLICING_FINALLY_CLEAN(1);
/* OK, we have the isoforms, now we need the read positions,
these are uniformly sampled from the individual isoforms. */
SPLICING_CHECK(splicing_vector_int_resize(position, noreads));
SPLICING_CHECK(splicing_vector_int_init(&exstart, 0));
SPLICING_FINALLY(splicing_vector_int_destroy, &exstart);
SPLICING_CHECK(splicing_vector_int_init(&exend, 0));
SPLICING_FINALLY(splicing_vector_int_destroy, &exend);
SPLICING_CHECK(splicing_vector_int_init(&exidx, 0));
SPLICING_FINALLY(splicing_vector_int_destroy, &exidx);
SPLICING_CHECK(splicing_gff_exon_start_end(gff, &exstart, &exend, &exidx,
gene));
/* Positions in isoform coordinates first */
for (i=0; i<noreads; i++) {
int iso=VECTOR(*isoform)[i];
int len=VECTOR(effisolen)[iso];
VECTOR(*position)[i]=RNG_INTEGER(1, len);
}
/* Translate isoform coordinates to genomic coordintes */
/* TODO: some of this is already calculated */
SPLICING_CHECK(splicing_iso_to_genomic(gff, gene, isoform, /*converter=*/ 0,
position));
/* CIGAR strings */
splicing_strvector_clear(cigar);
SPLICING_CHECK(splicing_strvector_reserve(cigar, noreads));
for (i=0; i<noreads; i++) {
char tmp[1000], *tmp2=tmp;
int iso=VECTOR(*isoform)[i];
size_t rs=VECTOR(*position)[i];
int ex=0;
int rl=readLength;
for (ex=VECTOR(exidx)[iso]; VECTOR(exend)[ex] < rs; ex++) ;
while (VECTOR(exend)[ex] < rs+rl-1) {
tmp2 += snprintf(tmp2, sizeof(tmp)/sizeof(char)-(tmp2-tmp)-1, "%iM%iN",
(int) (VECTOR(exend)[ex]-rs+1),
(int) (VECTOR(exstart)[ex+1]-VECTOR(exend)[ex]-1));
if (tmp2 >= tmp + sizeof(tmp)/sizeof(char)) {
SPLICING_ERROR("CIGAR string too long", SPLICING_EINVAL);
}
rl -= (VECTOR(exend)[ex] - rs + 1);
rs = VECTOR(exstart)[ex+1];
ex++;
}
tmp2 += snprintf(tmp2, sizeof(tmp)/sizeof(char)-(tmp2-tmp)-1, "%iM", rl);
if (tmp2 >= tmp + sizeof(tmp)/sizeof(char)) {
SPLICING_ERROR("CIGAR string too long", SPLICING_EINVAL); }
SPLICING_CHECK(splicing_strvector_append(cigar, tmp));
}
splicing_vector_int_destroy(&exidx);
splicing_vector_int_destroy(&exend);
splicing_vector_int_destroy(&exstart);
splicing_vector_int_destroy(&effisolen);
SPLICING_FINALLY_CLEAN(4);
return 0;
}
int splicing_miso(const splicing_gff_t *gff, size_t gene,
const splicing_vector_int_t *position,
const char **cigarstr, int readLength,
int noIterations, int noBurnIn, int noLag,
const splicing_vector_t *hyperp,
splicing_matrix_t *samples, splicing_vector_t *logLik,
splicing_matrix_t *match_matrix,
splicing_matrix_t *class_templates,
splicing_vector_t *class_counts,
splicing_vector_int_t *assignment,
splicing_miso_rundata_t *rundata) {
double acceptP, cJS, pJS, sigma;
int noReads = splicing_vector_int_size(position);
splicing_vector_int_t *myass=assignment, vass;
size_t noiso;
splicing_vector_t vpsi, vpsiNew, valpha, valphaNew,
*psi=&vpsi, *psiNew=&vpsiNew, *alpha=&valpha, *alphaNew=&valphaNew;
int noSamples = (noIterations - noBurnIn + 1) / noLag;
int i, m, lagCounter=0, noS=0;
splicing_matrix_t *mymatch_matrix=match_matrix, vmatch_matrix;
splicing_vector_int_t match_order;
splicing_vector_int_t effisolen;
splicing_vector_t isoscores;
if ( (class_templates ? 1 : 0) + (class_counts ? 1 : 0) == 1) {
SPLICING_ERROR("Only one of `class_templates' and `class_counts' is "
"given", SPLICING_EINVAL);
}
SPLICING_CHECK(splicing_gff_noiso_one(gff, gene, &noiso));
rundata->noIso=noiso;
rundata->noIters=noIterations;
rundata->noBurnIn=noBurnIn;
rundata->noLag=noLag;
rundata->noAccepted = rundata->noRejected = 0;
if (assignment) {
SPLICING_CHECK(splicing_vector_int_resize(myass, noReads));
splicing_vector_int_null(myass);
} else {
myass=&vass;
SPLICING_CHECK(splicing_vector_int_init(myass, noReads));
SPLICING_FINALLY(splicing_vector_int_destroy, myass);
}
SPLICING_CHECK(splicing_vector_init(&vpsi, noiso));
SPLICING_FINALLY(splicing_vector_destroy, &vpsi);
SPLICING_CHECK(splicing_vector_init(&vpsiNew, noiso));
SPLICING_FINALLY(splicing_vector_destroy, &vpsiNew);
SPLICING_CHECK(splicing_vector_init(&valpha, noiso-1));
SPLICING_FINALLY(splicing_vector_destroy, &valpha);
SPLICING_CHECK(splicing_vector_init(&valphaNew, noiso-1));
SPLICING_FINALLY(splicing_vector_destroy, &valphaNew);
if (match_matrix) {
SPLICING_CHECK(splicing_matrix_resize(match_matrix, noiso, noReads));
} else {
mymatch_matrix=&vmatch_matrix;
SPLICING_CHECK(splicing_matrix_init(mymatch_matrix, noiso, noReads));
SPLICING_FINALLY(splicing_matrix_destroy, mymatch_matrix);
}
SPLICING_CHECK(splicing_vector_int_init(&match_order, noReads));
SPLICING_FINALLY(splicing_vector_int_destroy, &match_order);
SPLICING_CHECK(splicing_matchIso(gff, gene, position, cigarstr,
mymatch_matrix));
SPLICING_CHECK(splicing_order_matches(mymatch_matrix, &match_order));
if (class_templates && class_counts) {
SPLICING_CHECK(splicing_i_miso_classes(mymatch_matrix, &match_order,
class_templates, class_counts,
/*bin_class_templates=*/ 0,
/*bin_class_counts=*/ 0));
}
SPLICING_CHECK(splicing_vector_int_init(&effisolen, noiso));
SPLICING_FINALLY(splicing_vector_int_destroy, &effisolen);
SPLICING_CHECK(splicing_vector_init(&isoscores, noiso));
SPLICING_FINALLY(splicing_vector_destroy, &isoscores);
SPLICING_CHECK(splicing_gff_isolength_one(gff, gene, &effisolen));
for (i=0; i<noiso; i++) {
int l=VECTOR(effisolen)[i]-readLength+1;
VECTOR(effisolen)[i] = l > 0 ? l : 0;
VECTOR(isoscores)[i] = -log((double) l);
}
SPLICING_CHECK(splicing_matrix_resize(samples, noiso, noSamples));
SPLICING_CHECK(splicing_vector_resize(logLik, noSamples));
/* Initialize Psi(0) randomly */
SPLICING_CHECK(splicing_drift_proposal(/* mode= */ 0, 0, 0, 0, 0, 0,
noiso, psi, alpha, &sigma, 0));
SPLICING_CHECK(splicing_drift_proposal(/* mode= */ 1, psi, alpha, sigma,
0, 0, noiso, psi, alpha, 0, 0));
/* Initialize assignments of reads */
SPLICING_CHECK(splicing_reassign_samples(mymatch_matrix, &match_order, psi,
noiso, myass));
/* foreach Iteration m=1, ..., M do */
for (m=0; m < noIterations; m++) {
SPLICING_CHECK(splicing_drift_proposal(/* mode= */ 1, psi, alpha, sigma,
0, 0, noiso, psiNew, alphaNew, 0,
0));
SPLICING_CHECK(splicing_metropolis_hastings_ratio(myass, noReads, psiNew,
alphaNew, psi, alpha,
sigma, noiso,
&effisolen, hyperp,
&isoscores,
m > 0 ? 1 : 0,
&acceptP, &cJS, &pJS));
if (acceptP >= 1 || RNG_UNIF01() < acceptP) {
splicing_vector_t *tmp;
tmp=psi; psi=psiNew; psiNew=tmp;
tmp=alpha; alpha=alphaNew; alphaNew=tmp;
cJS = pJS;
rundata->noAccepted ++;
} else {
rundata->noRejected ++;
}
if (m >= noBurnIn) {
if (lagCounter == noLag - 1) {
memcpy(&MATRIX(*samples, 0, noS), VECTOR(*psi),
noiso * sizeof(double));
VECTOR(*logLik)[noS] = cJS;
noS++;
lagCounter = 0;
} else {
lagCounter ++;
}
}
SPLICING_CHECK(splicing_reassign_samples(mymatch_matrix, &match_order,
psi, noiso, myass));
} /* for m < noIterations */
splicing_vector_destroy(&isoscores);
splicing_vector_int_destroy(&effisolen);
splicing_vector_int_destroy(&match_order);
SPLICING_FINALLY_CLEAN(3);
if (!match_matrix) {
splicing_matrix_destroy(mymatch_matrix);
SPLICING_FINALLY_CLEAN(1);
}
splicing_vector_destroy(&valphaNew);
splicing_vector_destroy(&valpha);
splicing_vector_destroy(&vpsiNew);
splicing_vector_destroy(&vpsi);
SPLICING_FINALLY_CLEAN(4);
if (!assignment) {
splicing_vector_int_destroy(myass);
SPLICING_FINALLY_CLEAN(1);
}
return 0;
}
int splicing_simulate_paired_reads(const splicing_gff_t *gff, int gene,
const splicing_vector_t *expression,
int noreads, int readLength,
const splicing_vector_t *fragmentProb,
int fragmentStart, double normalMean,
double normalVar, double numDevs,
splicing_vector_int_t *isoform,
splicing_vector_int_t *position,
splicing_strvector_t *cigar,
splicing_vector_t *sampleprob) {
size_t i, j, noiso, il, nogenes;
splicing_vector_t *mysampleprob=sampleprob, vsampleprob;
splicing_vector_t px, cpx;
double sumpx, sumpsi=0.0;
splicing_vector_int_t isolen;
int goodiso=0;
splicing_vector_int_t exstart, exend, exidx;
splicing_vector_t *myfragmentProb=(splicing_vector_t*) fragmentProb,
vfragmentProb;
int fs, fl;
SPLICING_CHECK(splicing_gff_nogenes(gff, &nogenes));
if (gene < 0 || gene >= nogenes) {
SPLICING_ERROR("Invalid gene id", SPLICING_EINVAL);
}
/* TODO: more error checks */
if (!fragmentProb) {
myfragmentProb=&vfragmentProb;
SPLICING_CHECK(splicing_vector_init(&vfragmentProb, 0));
SPLICING_FINALLY(splicing_vector_destroy, &vfragmentProb);
SPLICING_CHECK(splicing_normal_fragment(normalMean, normalVar, numDevs,
readLength, myfragmentProb,
&fragmentStart));
splicing_vector_scale(myfragmentProb,
1.0/splicing_vector_sum(myfragmentProb));
}
il=splicing_vector_size(myfragmentProb);
fs=fragmentStart;
fl=fragmentStart+il-1;
SPLICING_CHECK(splicing_gff_noiso_one(gff, gene, &noiso));
if ( fabs(splicing_vector_sum(myfragmentProb) - 1.0) > 1e-10 ) {
SPLICING_ERROR("Fragment length distribution does not sum up to 1",
SPLICING_EINVAL);
}
SPLICING_CHECK(splicing_vector_int_init(&isolen, noiso));
SPLICING_FINALLY(splicing_vector_int_destroy, &isolen);
SPLICING_CHECK(splicing_gff_isolength_one(gff, gene, &isolen));
SPLICING_CHECK(splicing_vector_copy(&px, myfragmentProb));
SPLICING_FINALLY(splicing_vector_destroy, &px);
SPLICING_CHECK(splicing_vector_init(&cpx, il));
SPLICING_FINALLY(splicing_vector_destroy, &cpx);
if (!sampleprob) {
mysampleprob=&vsampleprob;
SPLICING_CHECK(splicing_vector_init(mysampleprob, noiso));
SPLICING_FINALLY(splicing_vector_destroy, mysampleprob);
} else {
SPLICING_CHECK(splicing_vector_resize(mysampleprob, noiso));
}
for (sumpx=VECTOR(px)[0], i=1; i<il; i++) {
VECTOR(px)[i] += VECTOR(px)[i-1];
sumpx += VECTOR(px)[i];
}
VECTOR(cpx)[0] = VECTOR(px)[0];
for (i=1; i<il; i++) {
VECTOR(cpx)[i] = VECTOR(cpx)[i-1] + VECTOR(px)[i];
}
for (i=0; i<noiso; i++) {
int ilen=VECTOR(isolen)[i];
int r1= ilen >= fl ? ilen - fl + 1 : 0;
int r2= ilen >= fs ? (ilen >= fl ? fl - fs : ilen - fs + 1) : 0;
/* int r3= fs - 1; */
double sp=0.0;
if (r1 > 0) { sp += r1; }
if (r2 > 0) { sp += VECTOR(cpx)[r2-1]; }
VECTOR(*mysampleprob)[i] = sp * VECTOR(*expression)[i];
if (VECTOR(*mysampleprob)[i] != 0) { goodiso += 1; }
sumpsi += VECTOR(*mysampleprob)[i];
}
if (goodiso == 0) {
SPLICING_ERROR("No isoform is possible", SPLICING_FAILURE);
}
for (i=1; i<noiso; i++) {
VECTOR(*mysampleprob)[i] += VECTOR(*mysampleprob)[i-1];
}
SPLICING_CHECK(splicing_vector_int_resize(isoform, noreads*2));
for (i=0; i<2*noreads; i+=2) {
int w;
double rand;
if (noiso==1) {
w=0;
} else if (noiso==2) {
rand = RNG_UNIF01() * sumpsi;
w = (rand < VECTOR(*mysampleprob)[0]) ? 0 : 1;
} else {
rand = RNG_UNIF01() * sumpsi;
for (w=0; rand > VECTOR(*mysampleprob)[w]; w++) ;
}
VECTOR(*isoform)[i]=VECTOR(*isoform)[i+1]=w;
}
if (!sampleprob) {
splicing_vector_destroy(mysampleprob);
SPLICING_FINALLY_CLEAN(1);
} else {
for (i=noiso-1; i>0; i--) {
VECTOR(*mysampleprob)[i] -= VECTOR(*mysampleprob)[i-1];
}
}
/* We have the isoforms, now get the read positions. */
SPLICING_CHECK(splicing_vector_int_resize(position, noreads*2));
SPLICING_CHECK(splicing_vector_int_init(&exstart, 0));
SPLICING_FINALLY(splicing_vector_int_destroy, &exstart);
SPLICING_CHECK(splicing_vector_int_init(&exend, 0));
SPLICING_FINALLY(splicing_vector_int_destroy, &exend);
SPLICING_CHECK(splicing_vector_int_init(&exidx, 0));
SPLICING_FINALLY(splicing_vector_int_destroy, &exidx);
SPLICING_CHECK(splicing_gff_exon_start_end(gff, &exstart, &exend, &exidx,
gene));
/* Positions in isoform coordinates first.
These are sampled based on the fragment length distribution. */
for (i=0, j=0; i<noreads; i++) {
int iso=VECTOR(*isoform)[2*i];
int ilen=VECTOR(isolen)[iso];
int r1= ilen >= fl ? ilen - fl + 1 : 0;
int r2= ilen >= fs ? (ilen >= fl ? fl - fs : ilen - fs + 1) : 0;
/* int r3= fs - 1; */
int pos, fragment;
double sp=0.0;
if (r1 > 0) { sp += r1; }
if (r2 > 0) { sp += VECTOR(cpx)[r2-1]; }
double rand=RNG_UNIF(0, sp);
if (rand < r1) {
pos = ceil(rand);
} else {
int w;
rand -= r1;
for (w=0; VECTOR(cpx)[w] < rand; w++) ;
pos = r1 + r2 - w;
}
if (pos <= r1) {
rand=RNG_UNIF(0, 1.0);
} else {
rand=RNG_UNIF(0, VECTOR(px)[r1+r2-pos]);
}
for (fragment=0; VECTOR(px)[fragment] < rand; fragment++) ;
fragment += fragmentStart;
VECTOR(*position)[j++] = pos;
VECTOR(*position)[j++] = pos+fragment-readLength;
}
/* Translate positions to genomic coordinates */
/* TODO: some of this is already calculated */
SPLICING_CHECK(splicing_iso_to_genomic(gff, gene, isoform, /*converter=*/ 0,
position));
/* CIGAR strings */
splicing_strvector_clear(cigar);
SPLICING_CHECK(splicing_strvector_reserve(cigar, 2*noreads));
for (j=0; j<2*noreads; j++) {
char tmp[1000], *tmp2=tmp;
int iso=VECTOR(*isoform)[j];
size_t rs=VECTOR(*position)[j];
int ex=0;
int rl=readLength;
for (ex=VECTOR(exidx)[iso]; VECTOR(exend)[ex] < rs; ex++) ;
while (rs + rl - 1 > VECTOR(exend)[ex]) {
tmp2 += snprintf(tmp2, sizeof(tmp)/sizeof(char)-(tmp2-tmp)-1, "%iM%iN",
(int) (VECTOR(exend)[ex]-rs+1),
(int) (VECTOR(exstart)[ex+1]-VECTOR(exend)[ex]-1));
if (tmp2 >= tmp + sizeof(tmp)/sizeof(char)) {
SPLICING_ERROR("CIGAR string too long", SPLICING_EINVAL);
}
rl -= (VECTOR(exend)[ex] - rs + 1);
rs = VECTOR(exstart)[ex+1];
ex++;
}
tmp2 += snprintf(tmp2, sizeof(tmp)/sizeof(char)-(tmp2-tmp)-1, "%iM", rl);
if (tmp2 >= tmp + sizeof(tmp)/sizeof(char)) {
SPLICING_ERROR("CIGAR string too long", SPLICING_EINVAL);
}
SPLICING_CHECK(splicing_strvector_append(cigar, tmp));
}
splicing_vector_int_destroy(&exidx);
splicing_vector_int_destroy(&exend);
splicing_vector_int_destroy(&exstart);
splicing_vector_destroy(&cpx);
splicing_vector_destroy(&px);
splicing_vector_int_destroy(&isolen);
SPLICING_FINALLY_CLEAN(6);
if (!fragmentProb) {
splicing_vector_destroy(myfragmentProb);
SPLICING_FINALLY_CLEAN(1);
}
return 0;
}