GUHINDUKA
kamere
GUSHYIKIRANA
Inyandiko-mvugo yuzuye iranga SF3B1 ihindagurika muri lymphocytike leukemia idakira igaragaza kugabanuka kwimitsi yagumishijwe
Inyandiko-mvugo yuzuye |Urukurikirane rwa Nanopore |Ubundi isesengura rya isoform
Amavu n'amavuko
SGuhinduranya kwa omatic muburyo bwo gutera SF3B1 byavuzwe cyane ko bifitanye isano na kanseri zitandukanye, harimo na lymphocytike leukemia idakira (CLL), uveal melanoma, kanseri y'ibere, n'ibindi.Nyamara, ubushakashatsi kuri ubwo buryo butandukanye bwo gutondeka bwagiye bugarukira gusa ku byabaye-urwego no kubura ubumenyi kurwego rwa isoform bitewe no kugabanya inyandiko-mvugo isomwe.Hano, urubuga rwa nanopore rwatangijwe kugirango rutange inyandiko-mvugo yuzuye, yahaye imbaraga inverstigation kuri AS isoforms.
Igishushanyo mbonera
Ubushakashatsi
Itsinda:1. CLL-SF3B1 (WT) 2. CLL-SF3B1 (mutation ya K700E);3. B-selile isanzwe
Ingamba zikurikirana:Minion 2D ikurikirana isomero, PromethION 1D ikurikirana isomero;bigufi-soma amakuru kuva kurugero rumwe
Urubuga rukurikirana:UMUNTU WA MBERE;Isezerano rimwe;
Isesengura ryibinyabuzima
Ibisubizo
A.yose hamwe yasomwe miliyoni 257 yavuye muri 6 CLL hamwe na 3 B-selile.Ugereranije, 30.5% by'ibi bisomwa byagaragaye nk'inyandiko-mvugo yuzuye.
Full-uburebure ubundi isoform isesengura rya RNA (FLAIR) ryakozwe kugirango habeho urwego rwicyizere cyinshi isoforms.FLAIR irashobora kuvugwa muri make nka:
Nanopore asoma guhuza: menya imiterere rusange yinyandiko-mvugo ishingiye kuri genome;
SIgice cyo guhuza gukosora: gukosora amakosa akurikirana (umutuku) hamwe na site igabanyijemo ibice byombi bisobanuwe, inyuguti ziva mubisomwa bigufi cyangwa byombi;
Collapse: vuga muri make isoforms ihagarariye ishingiye kumacakubiri ihuza (umurongo-wambere).Hitamo ibyiringiro byinshi isofrom ukurikije umubare wogushigikira gusoma (Threshold: 3).
Igicapo 1. Isesengura RIKURIKIRA kugirango umenye isoforms yuzuye ijyanye na mutation ya SF3B1 muri CLL
FLAIR Yamenyekanye 326,699-yizeye cyane yatanzwe isoforms, 90% muri zo ni isoforms.Byinshi muribi isoforms itamenyekanye wasangaga aribintu bishya byerekana ibice bizwi (142.971), mugihe isoforms isigaye yarimo intron yagumanye (21,700) cyangwa igitabo cyitwa exon (3594).
Long-soma ikurikiranye itanga imbaraga zo kumenya mutant SF3B1-K700E -guhindura ibice byimbuga kurwego rwa isoform.35 ubundi buryo bwa 3'SS na 10 ubundi 5'SS byagaragaye ko bitandukanye cyane hagati ya SF3B1-K700E na SF3B1-WT.33 kuri 35 byahinduwe byavumbuwe nuburyo bukurikiranye gusoma.Mumibare ya Nanopore, ikwirakwizwa ryintera hagati ya SF3B1-K700E-yahinduwe 3'SS kurubuga rwa Canonical peaks ni -20 bp, itandukanye cyane no gukwirakwiza igenzura, bisa nibyavuzwe muri CLL ikurikiranye igihe gito.Isoforms ya gene ya ERGIC3 yasesenguwe, aho isoform yigitabo irimo urubuga rwacitsemo ibice wasangaga ari nyinshi muri SF3B1-K700E.Byombi byegeranye kandi byitaruye 3'SS byari bifitanye isano na AS itandukanye itanga isoforms nyinshi.
Igishushanyo 2. Ubundi buryo 3 ′ uburyo bwo gutondeka bwerekanwe na nanopore ikurikirana
Isesengura ry'imikoreshereze ya IR ryabaye ndende mugihe gito-gisomwe gishingiye kubisesengura bitewe nicyizere cyo kumenya IR no kugereranya.Kugaragaza isoforms ya IR muri SF3B1-K700E na SF3B1-WT byapimwe hashingiwe ku ruhererekane rwa nanopore, byerekana ko isi yose igenga isoforms muri SF3B1-K700E.
Igicapo 4. Ubwinshi bwubuhinzi no guhuza imiyoboro muburyo butatu bwo guhinga (A na B);Isesengura ryamashyamba risanzwe (C) nubusabane hagati yubuhinzi nubukoloni bwa AMF (D)
Igicapo 3. Ibikorwa byo gukodesha imbere biragabanijwe cyane muri CLL SF3B1-K700E
Ikoranabuhanga
Nanopore Gusoma igihe kirekire
NGukurikirana kwa anopore ni molekile imwe nyayo-nyayo ya tekinoroji ikurikirana.
DADN cyangwa RNA ihujwe na poroteyine ya nanoporus yashyizwe muri biofilm kandi itabishaka iyobowe na poroteyine.
DImigozi ya NA / RNA inyura kuri nanopore umuyoboro wa poroteyine ku gipimo runaka ukurikije itandukaniro rya voltage.
Molecules itanga ibimenyetso byamashanyarazi bitandukanye ukurikije imiterere yimiti.
Real-igihe cyo gutondekanya ibyagezweho bigerwaho no guhamagarwa shingiro.
Imikorere yuburebure bwuzuye bwuzuye
S Kwuzuza amakuru
Inshuro 7 zasomwe zisabwa kugirango ugere kubintu byuzuye.
Ident Kumenyekanisha Imiterere
Kumenyekanisha ibintu bitandukanye byubatswe hamwe byumvikanyweho byuzuye-gusoma buri nyandiko-mvugo
Analyse Inyandiko-mvugo itandukanye isesengura -Guhishura impinduka zihishwa-bigufi
Reba
Tang AD, Soulette CM, Baren MJV, nibindi.Inyandiko-mvugo yuzuye iranga ihinduka rya SF3B1 muri lymphocytike ya leukemiya idakira igaragaza kugabanuka kwimitsi yagumishijwe [J].Itumanaho rya Kamere.
Ikoranabuhanga hamwe nibyingenzi igamije gusangira ibyagezweho vuba muburyo butandukanye bwo gukoresha tekinoroji ikurikirana muburyo butandukanye hamwe nibitekerezo byiza mugushushanya no gucukura amakuru.
Igihe cyo kohereza: Mutarama-08-2022