TRANSCRIPTOMICS
yanayi
SADARWA
Cikakkun bayanan kwafi na maye gurbi na SF3B1 a cikin cutar sankarar lymphocytic na yau da kullun yana bayyana raguwar tsarin intron da aka riƙe.
Cikakkun bayanai |Tsarin Nanopore|Madadin nazarin isoform
Fage
Somatic maye gurbi a cikin splicing factor SF3B1 an ko'ina ya yi tarayya da cututtuka daban-daban, ciki har da na kullum lymphocytic cutar sankarar bargo (CLL), uveal melanoma, nono ciwon daji, da dai sauransu Bugu da kari, short-karanta transcriptomic karatu sun bayyana aberrant splicing alamu jawo SF3B1 maye gurbi.Duk da haka, nazarin kan waɗannan madadin tsarin rarrabawa ya daɗe yana iyakance ga matakin taron da kuma rashin ilimi akan matakin isoform saboda taƙaitaccen taƙaitaccen rubutun da aka haɗa.Anan, an gabatar da dandamali na jerin nanopore don samar da cikakkun bayanai na tsawon lokaci, wanda ya ba da ikon bincike akan isoforms AS.
Tsarin Gwaji
Gwaje-gwaje
Rukuni:1. CLL-SF3B1 (WT) 2. CLL-SF3B1 (K700E maye gurbi);3. Kwayoyin B na al'ada
Dabarun tsarawa:MinION 2D jerin laburare, PromethION 1D jerin laburare;gajeriyar karanta bayanai daga samfurori iri ɗaya
Dandalin jerin abubuwa:ABIN MINION;ONT PromethION;
Bioinformatic Analysis
Sakamako
AAn samar da adadin karatun miliyan 257 daga samfuran CLL 6 da ƙwayoyin B-3.A matsakaita na 30.5% na waɗannan karantawa an gano su azaman cikakkun bayanai.
FCanjin canjin isoform na RNA(FLAIR) mai tsayi mai tsayi an haɓaka shi don samar da saiti na isoforms masu ƙarfi.Ana iya taƙaita FLAIR kamar:
Nanopore yana karanta jeri: gano tsarin kwafi na gabaɗaya dangane da kwayoyin halitta;
Sgyara junction plice: daidai kurakuran jeri (ja) tare da rukunin yanar gizo daga ko dai annotate introns, introns daga gajerun bayanan karantawa ko duka biyu;
Collapse: taƙaita isoforms na wakilci bisa ga sarƙoƙin haɗin gwiwa (saitin farko na farko).Zaɓi isofrom babban ƙarfin gwiwa dangane da adadin karantawa mai goyan baya (Mafafin: 3).
Hoto 1. Binciken FLAIR don gano cikakken isoforms masu tsayi da ke hade da maye gurbin SF3B1 a cikin CLL
FLAIR An Gano 326,699 babban amintaccen tsattsauran ra'ayi, 90% na waɗancan sifofi ne na zamani.Yawancin waɗannan isoforms waɗanda ba a bayyana su ba an samo su ne haɗuwar sabbin abubuwan haɗin gwiwa na sanannun splice juntions (142,971), yayin da sauran isoforms na novel sun ƙunshi ko dai riƙe intron (21,700) ko novel exon (3594).
Ljeri-karanta ong yana ba da ikon gano mutant SF3B1-K700E -canza rukunin rukunin yanar gizo a matakin isoform.35 madadin 3'SSs da 10 madadin 5'SSs an gano suna da banbanci sosai tsakanin SF3B1-K700E da SF3B1-WT.33 daga cikin 35 canje-canje an gano su ta hanyar jerin dogon karatu.A cikin bayanan Nanopore, rarraba tazara tsakanin SF3B1-K700E-canza 3'SSs zuwa ga wuraren canonical kololuwa yana kusa da -20 bp, wanda ya bambanta sosai da rarrabawar sarrafawa, kama da abin da aka ruwaito a cikin jerin gajeren karanta CLL.Isoforms na ERGIC3 gene an bincikar su, inda aka sami isoform na labari wanda ke dauke da wurin da ake kusa da shi a cikin SF3B1-K700E.Dukansu na kusa da 3'SS na nesa suna da alaƙa da keɓaɓɓen tsarin AS wanda ke haifar da isoforms da yawa.
Hoto 2. Madadin 3′ tsarin rarrabawa da aka gano tare da bayanan jeri na nanopore
Binciken amfani da taron na IR ya daɗe yana iyakancewa a cikin taƙaitaccen bincike na tushen karantawa saboda amincewa ga gano IR da ƙididdigewa.Ma'anar isoforms na IR a cikin SF3B1-K700E da SF3B1-WT an ƙididdige su bisa ga jerin nanopore, yana nuna ƙa'ida ta duniya na IR isoforms a cikin SF3B1-K700E.
Hoto 4. Ƙarfin aikin noma da haɗin yanar gizo a cikin tsarin noma guda uku (A da B);Binciken gandun daji na bazuwar (C) da alaƙa tsakanin ƙarfin aikin gona da mulkin mallaka na AMF (D)
Hoto 3. Abubuwan riƙewa na gaba sun fi ƙasƙantar da su a cikin CLL SF3B1-K700E
Fasaha
Tsarin Nanopore Tsawon Karatu
Njerin anopore fasaha ce ta siginar siginar guda ɗaya na ainihin lokacin.
DDNA ko RNA mai ɗaure kai zai ɗaure da furotin nanoporous da aka saka a cikin biofilm da kwancewa ƙarƙashin jagorancin furotin mota.
DMatsalolin NA/RNA suna wucewa ta hanyar furotin na nanopore a wani ƙayyadadden ƙimar ƙarƙashin aikin bambancin wutar lantarki.
Molecules suna haifar da siginar lantarki daban-daban bisa ga tsarin sinadarai.
RAna samun gano jerin jeri ta hanyar kiran tushe.
Aiwatar da jeri mai cikakken tsayin rubutu
√ Cikewar Data
Ana buƙatar ƙarancin karantawa mai ninki 7 don isa daidaitattun bayanai.
√ Ƙididdigar Tsarin Rubutu
Gano bambance-bambancen tsari daban-daban tare da cikakken cikakken karantawa na kowane kwafin.
√ Nazari na banbance-banbance-matakin kwafi-Bayyana canje-canjen da aka ɓoye ta gajerun karantawa
Magana
Tang AD , Soulette CM , Baren MJV , et al.Cikakkun bayanan kwafi na SF3B1 maye gurbi a cikin cutar sankarar lymphocytic na yau da kullun yana bayyana raguwar abubuwan da aka riƙe [J].Sadarwar yanayi.
Fasaha da Manyan Labarai yana da nufin raba mafi kyawun aikace-aikacen nasara na baya-bayan nan na fasahohi daban-daban na babban abin da ake aiwatarwa a fagen bincike daban-daban da kuma ƙwararrun dabaru a ƙirar gwaji da hakar bayanai.
Lokacin aikawa: Janairu-08-2022