Simple Walkthrough

Overview

The purpose of this walkthrough will be to illustrate a simple, but complete example of using gNALI to find high-confidence loss-of-function variants of genes.

Input Data

The input file should contain a list of genes as HGNC symbols, separated by a newline character. Genes in non-standard formats will not be analyzed.

The example input file located in the following location:

inputs/genes.txt

The example input file contains the following contents and can be found here:

> cat inputs/genes.txt
CCR5
ALCAM

Running gNALI

gNALI only requires an input file, and has other optional parameters. We will use an input file with the path inputs/genes.txt, filter for high-confidence loss-of-function variants, and write our results to a directory called output-simple.

Since we have not selected a database to use, gNALI will default to gnomADv2.1.1.

Here is what such a command would look like:

gnali
    --input inputs/genes.txt
    --output output-simple/

Output

By default, gNALI will have two output files in output-simple/: a basic output file, and a detailed output file. When using the -v/--vcf flag, a third additional output file will be generated. An example of such output can be found in the advanced walkthrough. More information on outputs can be found here.

Output files for this example can be found here.

Basic Output

The basic output file contains a subset of the input genes, the ones that have high-confidence loss-of-function variants that pass filtering. It also contains a list of genes that could not be found in the specified database, if any.

The file shown below can also be found here.

output-simple/Nonessential_Host_Genes_\(Basic\).txt:
HGNC_Symbol Status
CCR5 HC LoF found
ALCAM HC LoF found

Detailed Output

The detailed output file contains the high-confidence loss-of-function variants that pass filtering with some annotations extracted.

The file shown below can also be found here.

output-simple/Nonessential_Host_Genes_\(Detailed\).txt:
Chromosome Position_Start RSID Reference_Allele Alternate_Allele Score Quality LoF_Variant LoF_Annotation HGNC_Symbol Ensembl_Code HGVSc
3 46414403 rs748244565 CAA C 2676.90 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.11_12delAA
3 46414403 rs748244565 CAA C 2676.90 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.11_12delAA
3 46414403 rs748244565 CAA C 2676.90 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.11_12delAA
3 46414434 rs758662716 AT A 4280.25 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.43delT
3 46414434 rs758662716 AT A 4280.25 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.43delT
3 46414434 rs758662716 AT A 4280.25 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.43delT
3 46414436 rs200209014 TA T 58824.12 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.44delA
3 46414436 rs200209014 TA T 58824.12 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.44delA
3 46414436 rs200209014 TA T 58824.12 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.44delA
3 46414443 rs369206494 C A 73505.83 PASS A stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.50C>A
3 46414443 rs369206494 C A 73505.83 PASS A stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.50C>A
3 46414443 rs369206494 C A 73505.83 PASS A stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.50C>A
3 46414453 rs760428955 C A 2656.37 PASS A stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.60C>A
3 46414453 rs760428955 C A 2656.37 PASS A stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.60C>A
3 46414453 rs760428955 C A 2656.37 PASS A stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.60C>A
3 46414454 rs780235820 C CA 3750.42 PASS A frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.67dupA
3 46414454 rs780235820 C CA 3750.42 PASS A frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.67dupA
3 46414454 rs780235820 C CA 3750.42 PASS A frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.67dupA
3 46414457 rs768398484 A T 2699.39 PASS T stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.64A>T
3 46414457 rs768398484 A T 2699.39 PASS T stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.64A>T
3 46414457 rs768398484 A T 2699.39 PASS T stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.64A>T
3 46414541 rs747388089 CT C 6002.15 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.149delT
3 46414541 rs747388089 CT C 6002.15 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.149delT
3 46414541 rs747388089 CT C 6002.15 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.149delT
3 46414604 rs769057798 AACCTGGC A 2763.36 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.212_218delACCTGGC
3 46414604 rs769057798 AACCTGGC A 2763.36 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.212_218delACCTGGC
3 46414604 rs769057798 AACCTGGC A 2763.36 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.212_218delACCTGGC
3 46414651 rs777330502 G A 25606.70 PASS A stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.258G>A
3 46414651 rs777330502 G A 25606.70 PASS A stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.258G>A
3 46414651 rs777330502 G A 25606.70 PASS A stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.258G>A
3 46414696 rs1800560 T A 540878.23 PASS A stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.303T>A
3 46414696 rs1800560 T A 540878.23 PASS A stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.303T>A
3 46414696 rs1800560 T A 540878.23 PASS A stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.303T>A
3 46414697 rs1294651548 C T 5088.14 PASS T stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.304C>T
3 46414697 rs1294651548 C T 5088.14 PASS T stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.304C>T
3 46414697 rs1294651548 C T 5088.14 PASS T stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.304C>T
3 46414702 rs56068070 CTT C 19747.17 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.310_311delTT
3 46414702 rs56068070 CTT C 19747.17 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.310_311delTT
3 46414702 rs56068070 CTT C 19747.17 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.310_311delTT
3 46414714 rs1249868905 C CTA 1526.36 PASS TA frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.323_324dupAT
3 46414714 rs1249868905 C CTA 1526.36 PASS TA frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.323_324dupAT
3 46414714 rs1249868905 C CTA 1526.36 PASS TA frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.323_324dupAT
3 46414774 rs1372217574 C G 950.36 PASS G stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.381C>G
3 46414774 rs1372217574 C G 950.36 PASS G stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.381C>G
3 46414774 rs1372217574 C G 950.36 PASS G stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.381C>G
3 46414791 rs1215591945 CTG C 1355.36 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.402_403delGT
3 46414791 rs1215591945 CTG C 1355.36 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.402_403delGT
3 46414791 rs1215591945 CTG C 1355.36 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.402_403delGT
3 46414909 rs1188964664 AG A 712.37 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.518delG
3 46414909 rs1188964664 AG A 712.37 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.518delG
3 46414909 rs1188964664 AG A 712.37 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.518delG
3 46414935 rs938517991 AT A 9974.16 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.546delT
3 46414935 rs938517991 AT A 9974.16 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.546delT
3 46414935 rs938517991 AT A 9974.16 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.546delT
3 46414943 rs775750898 TACAGTCAGTATCAATTCTGGAAGAATTTCCAG T 74264261.52 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.554_585delGTCAGTATCAATTCTGGAAGAATTTCCAGACA
3 46414943 rs775750898 TACAGTCAGTATCAATTCTGGAAGAATTTCCAG T 74264261.52 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.554_585delGTCAGTATCAATTCTGGAAGAATTTCCAGACA
3 46414943 rs775750898 TACAGTCAGTATCAATTCTGGAAGAATTTCCAG T 74264261.52 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.554_585delGTCAGTATCAATTCTGGAAGAATTTCCAGACA
3 46414945 rs746626584 C G 74247720.02 PASS G stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.552C>G
3 46414945 rs746626584 C G 74247720.02 PASS G stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.552C>G
3 46414945 rs746626584 C G 74247720.02 PASS G stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.552C>G
3 46414949 rs113869679 C T 74238735.71 PASS T stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.556C>T
3 46414949 rs113869679 C T 74238735.71 PASS T stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.556C>T
3 46414949 rs113869679 C T 74238735.71 PASS T stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.556C>T
3 46415066 rs146972949 C T 120238.89 PASS T stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.673C>T
3 46415066 rs146972949 C T 120238.89 PASS T stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.673C>T
3 46415066 rs146972949 C T 120238.89 PASS T stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.673C>T
3 46415168 rs910924386 AC A 156.36 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.777delC
3 46415168 rs910924386 AC A 156.36 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.777delC
3 46415168 rs910924386 AC A 156.36 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.777delC
3 46415174 rs775289875 C T 8429.37 PASS T stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.781C>T
3 46415174 rs775289875 C T 8429.37 PASS T stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.781C>T
3 46415174 rs775289875 C T 8429.37 PASS T stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.781C>T
3 46415177 rs1446832405 G T 700.38 PASS T stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.784G>T
3 46415177 rs1446832405 G T 700.38 PASS T stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.784G>T
3 46415177 rs1446832405 G T 700.38 PASS T stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.784G>T
3 46415238 rs1294154353 CAG C 1387.53 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.849_850delGA
3 46415238 rs1294154353 CAG C 1387.53 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.849_850delGA
3 46415238 rs1294154353 CAG C 1387.53 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.849_850delGA
3 46415271 rs774050135 AC A 10605.94 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.882delC
3 46415271 rs774050135 AC A 10605.94 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.882delC
3 46415271 rs774050135 AC A 10605.94 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.882delC
3 46414434 rs758662716 AT A 296.46 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.43delT
3 46414434 rs758662716 AT A 296.46 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.43delT
3 46414434 rs758662716 AT A 296.46 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.43delT
3 46414696 rs1800560 T A 16687.73 PASS A stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.303T>A
3 46414696 rs1800560 T A 16687.73 PASS A stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.303T>A
3 46414696 rs1800560 T A 16687.73 PASS A stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.303T>A
3 46414943 rs775750898 TACAGTCAGTATCAATTCTGGAAGAATTTCCAG T 1947603.90 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000292303.4:c.554_585delGTCAGTATCAATTCTGGAAGAATTTCCAGACA
3 46414943 rs775750898 TACAGTCAGTATCAATTCTGGAAGAATTTCCAG T 1947603.90 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000343801.4:c.554_585delGTCAGTATCAATTCTGGAAGAATTTCCAGACA
3 46414943 rs775750898 TACAGTCAGTATCAATTCTGGAAGAATTTCCAG T 1947603.90 PASS - frameshift_variant CCR5 ENSG00000160791 ENST00000445772.1:c.554_585delGTCAGTATCAATTCTGGAAGAATTTCCAGACA
3 46414949 rs113869679 C T 1942568.40 PASS T stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.556C>T
3 46414949 rs113869679 C T 1942568.40 PASS T stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.556C>T
3 46414949 rs113869679 C T 1942568.40 PASS T stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.556C>T
3 46415066 rs146972949 C T 3449.38 PASS T stop_gained CCR5 ENSG00000160791 ENST00000292303.4:c.673C>T
3 46415066 rs146972949 C T 3449.38 PASS T stop_gained CCR5 ENSG00000160791 ENST00000343801.4:c.673C>T
3 46415066 rs146972949 C T 3449.38 PASS T stop_gained CCR5 ENSG00000160791 ENST00000445772.1:c.673C>T
3 105243261 rs1401656526 T TAATGC 3920.14 PASS AATGC frameshift_variant ALCAM ENSG00000170017 ENST00000306107.5:c.306_310dupTGCAA
3 105243261 rs1401656526 T TAATGC 3920.14 PASS AATGC frameshift_variant ALCAM ENSG00000170017 ENST00000472644.2:c.306_310dupTGCAA
3 105243261 rs1401656526 T TAATGC 3920.14 PASS AATGC frameshift_variant ALCAM ENSG00000170017 ENST00000486979.2:c.153_157dupTGCAA
3 105243324 rs748382272 TG T 3638.37 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000306107.5:c.367delG
3 105243324 rs748382272 TG T 3638.37 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000472644.2:c.367delG
3 105243324 rs748382272 TG T 3638.37 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000486979.2:c.214delG
3 105252448 rs763351683 TG T 904.36 PASS - frameshift_variant&splice_region_variant ALCAM ENSG00000170017 ENST00000306107.5:c.464delG
3 105252448 rs763351683 TG T 904.36 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000465413.2:c.6delG
3 105252448 rs763351683 TG T 904.36 PASS - frameshift_variant&splice_region_variant ALCAM ENSG00000170017 ENST00000472644.2:c.464delG
3 105252448 rs763351683 TG T 904.36 PASS - frameshift_variant&splice_region_variant ALCAM ENSG00000170017 ENST00000486979.2:c.311delG
3 105253574 rs753626110 GGAGTACAA G 26588.50 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000306107.5:c.618_625delGTACAAGA
3 105253574 rs753626110 GGAGTACAA G 26588.50 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000389927.4:c.45_52delGTACAAGA
3 105253574 rs753626110 GGAGTACAA G 26588.50 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000465413.2:c.160_167delGTACAAGA
3 105253574 rs753626110 GGAGTACAA G 26588.50 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000472644.2:c.618_625delGTACAAGA
3 105253574 rs753626110 GGAGTACAA G 26588.50 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000486979.2:c.465_472delGTACAAGA
3 105258923 rs1409380546 G T 230.36 PASS T stop_gained ALCAM ENSG00000170017 ENST00000306107.5:c.835G>T
3 105258923 rs1409380546 G T 230.36 PASS T stop_gained ALCAM ENSG00000170017 ENST00000472644.2:c.835G>T
3 105258923 rs1409380546 G T 230.36 PASS T stop_gained ALCAM ENSG00000170017 ENST00000486979.2:c.682G>T
3 105266064 rs748308492 C A 7063.46 PASS A stop_gained ALCAM ENSG00000170017 ENST00000306107.5:c.1176C>A
3 105266064 rs748308492 C A 7063.46 PASS A stop_gained ALCAM ENSG00000170017 ENST00000389927.4:c.342C>A
3 105266064 rs748308492 C A 7063.46 PASS A stop_gained ALCAM ENSG00000170017 ENST00000465413.2:c.457C>A
3 105266064 rs748308492 C A 7063.46 PASS A stop_gained ALCAM ENSG00000170017 ENST00000472644.2:c.1176C>A
3 105266064 rs748308492 C A 7063.46 PASS A stop_gained ALCAM ENSG00000170017 ENST00000486979.2:c.1023C>A
3 105266100 rs1189829981 AAG A 54.42 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000306107.5:c.1217_1218delAG
3 105266100 rs1189829981 AAG A 54.42 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000389927.4:c.383_384delAG
3 105266100 rs1189829981 AAG A 54.42 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000465413.2:c.498_499delAG
3 105266100 rs1189829981 AAG A 54.42 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000472644.2:c.1217_1218delAG
3 105266100 rs1189829981 AAG A 54.42 PASS - frameshift_variant ALCAM ENSG00000170017 ENST00000486979.2:c.1064_1065delAG
3 105266129 rs1165397359 G T 244.57 PASS T splice_donor_variant ALCAM ENSG00000170017 ENST00000306107.5:c.1240+1G>T
3 105266129 rs1165397359 G T 244.57 PASS T splice_donor_variant ALCAM ENSG00000170017 ENST00000389927.4:c.406+1G>T
3 105266129 rs1165397359 G T 244.57 PASS T splice_donor_variant ALCAM ENSG00000170017 ENST00000465413.2:c.521+1G>T
3 105266129 rs1165397359 G T 244.57 PASS T splice_donor_variant ALCAM ENSG00000170017 ENST00000472644.2:c.1240+1G>T
3 105266129 rs1165397359 G T 244.57 PASS T splice_donor_variant ALCAM ENSG00000170017 ENST00000486979.2:c.1087+1G>T
3 105268977 . G T 273.40 PASS T stop_gained ALCAM ENSG00000170017 ENST00000306107.5:c.1381G>T
3 105268977 . G T 273.40 PASS T stop_gained ALCAM ENSG00000170017 ENST00000389927.4:c.547G>T
3 105268977 . G T 273.40 PASS T stop_gained ALCAM ENSG00000170017 ENST00000465413.2:c.662G>T
3 105268977 . G T 273.40 PASS T stop_gained ALCAM ENSG00000170017 ENST00000472644.2:c.1381G>T
3 105268977 . G T 273.40 PASS T stop_gained ALCAM ENSG00000170017 ENST00000486979.2:c.1228G>T
3 105270986 rs1249936676 A G 151.43 PASS G splice_acceptor_variant ALCAM ENSG00000170017 ENST00000306107.5:c.1508-2A>G
3 105270986 rs1249936676 A G 151.43 PASS G splice_acceptor_variant ALCAM ENSG00000170017 ENST00000389927.4:c.674-2A>G
3 105270986 rs1249936676 A G 151.43 PASS G splice_acceptor_variant ALCAM ENSG00000170017 ENST00000465413.2:c.789-2A>G
3 105270986 rs1249936676 A G 151.43 PASS G splice_acceptor_variant ALCAM ENSG00000170017 ENST00000486979.2:c.1355-2A>G
3 105270987 rs764096754 G A 1329.70 PASS A splice_acceptor_variant ALCAM ENSG00000170017 ENST00000306107.5:c.1508-1G>A
3 105270987 rs764096754 G A 1329.70 PASS A splice_acceptor_variant ALCAM ENSG00000170017 ENST00000389927.4:c.674-1G>A
3 105270987 rs764096754 G A 1329.70 PASS A splice_acceptor_variant ALCAM ENSG00000170017 ENST00000465413.2:c.789-1G>A
3 105270987 rs764096754 G A 1329.70 PASS A splice_acceptor_variant ALCAM ENSG00000170017 ENST00000486979.2:c.1355-1G>A
3 105252445 rs1264516297 A T 208.47 PASS T splice_acceptor_variant ALCAM ENSG00000170017 ENST00000306107.5:c.460-2A>T
3 105252445 rs1264516297 A T 208.47 PASS T splice_acceptor_variant ALCAM ENSG00000170017 ENST00000472644.2:c.460-2A>T
3 105252445 rs1264516297 A T 208.47 PASS T splice_acceptor_variant ALCAM ENSG00000170017 ENST00000486979.2:c.307-2A>T