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Mkparịta ụka
Nkọwa ederede zuru oke nke SF3B1 mutation na leukemia lymphocytic na-adịghị ala ala na-ekpughe nhazi nke introns ejidere.
Ederede n'ogologo|Usoro Nanopore|Nyocha isoform ọzọ
ndabere
Smutations omatic in splicing factor SF3B1 ka a kọwapụtara n'ọtụtụ ebe na-ejikọta ya na ọrịa cancer dị iche iche, gụnyere ọrịa leukemia lymphocytic na-adịghị ala ala (CLL), melanoma uveal, ọrịa ara ara, wdg. Tụkwasị na nke a, ọmụmụ ihe odide ederede dị mkpirikpi ekpughere usoro nkwụsịtụ nke SF3B1 kpatara.Agbanyeghị, ọmụmụ gbasara usoro ngbanwe ndị a ka amachiri naanị na ọkwa mmemme yana enweghị ihe ọmụma na ọkwa isoform n'ihi mmachi nke ndebiri agụkọ gbakọtara.N'ebe a, ewebata usoro usoro nanopore iji mepụta transcript zuru oke, nke nyere ikike nyocha na isoforms AS.
Nleba anya
Nnwale
Ịchịkọta:1. CLL-SF3B1 (WT) 2. CLL-SF3B1 (K700E mutation);3. Nkịtị B-cell
Atụmatụ usoro:Usoro nke ọbá akwụkwọ MinION 2D, Usoro ọbá akwụkwọ PromethION 1D;mkpiri-agụ data si n'otu sample
Usoro nhazi usoro:ONT MINION;ONT PromethION;
Nyocha bioinformatic
Nsonaazụ
ANgụkọta nke nde 257 a na-agụ sitere na 6 CLL samples na 3 B-cell.Na nkezi, 30.5% nke ọgụgụ ndị a ka achọpụtara dị ka ndegharị n'ogologo.
FE mepụtara nyocha isoform ọzọ nke RNA(FLAIR) ogologo ogologo iji mepụta usoro nke isoforms nwere ntụkwasị obi dị elu.Enwere ike ichikota FLAIR dị ka:
Nanopore na-agụ nhazi: chọpụta nhazi ederede izugbe dabere na genome ntụaka;
Smgbazi plice junction: njehie usoro ziri ezi (acha ọbara ọbara) nwere saịtị splice sitere na introns nkọwapụta, introns sitere na data mkpirisi ọgụgụ ma ọ bụ ha abụọ;
Collapse: chịkọta ndị nnọchi anya isoforms dabere na splice junction chains (usoro ngafe mbụ).Họrọ isofrom nwere ntụkwasị obi dị elu dabere na ọnụ ọgụgụ agụọ na-akwado (Nkarịrị: 3).
Ọgụgụ 1. Nyocha FLAIR iji chọpụta isoforms zuru oke jikọtara ya na mutation SF3B1 na CLL.
FLAIR achọpụtara 326,699 nwere ntụkwasị obi dị elu, 90% nke bụ isoforms ọhụrụ.Achọpụtara ọtụtụ n'ime isoforms ndị a na-akọwaghị ka ọ bụ nchịkọta akwụkwọ akụkọ nke ama ama splice juntions (142,971), ebe isoforms akwụkwọ akụkọ ndị ọzọ nwere ma ọ bụ intron (21,700) ma ọ bụ akwụkwọ akụkọ exon (3594).
LUsoro ong-agụ na-enye ike njirimara nke mutant SF3B1-K700E-gbanwere saịtị splice na ọkwa isoform.Achọpụtara 35 ọzọ 3'SSs na 10 ọzọ 5'SS ka ekewapụrụ nke ukwuu n'etiti SF3B1-K700E na SF3B1-WT.33 n'ime 35 mgbanwe ka achọpụtara ọhụrụ site n'usoro ogologo ọgụgụ.Na data Nanopore, nkesa nkesa dị n'etiti SF3B1-K700E-gbanwere 3'SSs gaa na saịtị canonical dị gburugburu -20 bp, nke dị nnọọ iche na nkesa njikwa, dị ka nke a kọrọ na usoro CLL dị mkpirikpi.A na-enyocha mkpụrụ ndụ Isoforms nke mkpụrụ ndụ ERGIC3, ebe a chọtara isoform akwụkwọ akụkọ nwere saịtị splice dị nso karịa na SF3B1-K700E.Ejikọtara ma proximal na distal 3'SS na ụkpụrụ AS dị iche iche na-emepụta ọtụtụ isoforms.
Ọgụgụ 2. Nhọrọ 3′ ụkpụrụ splicing nke a chọpụtara na data usoro nke nanopore
Nyocha ojiji mmemme IR adịla ogologo oge na nyocha dabere na mkpirisi ọgụgụ n'ihi ntụkwasị obi na njirimara IR na nha.Ngosipụta nke isoforms IR na SF3B1-K700E na SF3B1-WT bụ ọnụ ọgụgụ dabere na usoro nanopore, na-ekpughe usoro nchịkwa zuru ụwa ọnụ nke isoforms IR na SF3B1-K700E.
Ọgụgụ 4. Ịrụ ọrụ ugbo na njikọ netwọk n'ofe usoro ugbo atọ (A na B);Nyocha oke ọhịa (C) na mmekọrịta dị n'etiti ike ọrụ ugbo na nchịkwa AMF (D)
Ọgụgụ 3. A na-achịkwa ihe omume njide intron nke ọma na CLL SF3B1-K700E
Nkà na ụzụ
Usoro Nanopore Ogologo ọgụgụ
Nsequencing anopore bụ otu molekul ezigbo oge usoro mgbama ọkụ eletrik.
DDNA ma ọ bụ RNA nwere ouble ga-ejikọta na protein nanoporous agbakwunyere na biofilm ma na-ewepụ n'okpuru ndu nke protein moto.
DỤdọ NA/RNA na-agafe na protein ọwa nanopore n'ogo ụfọdụ n'okpuru ọrụ nke ọdịiche voltaji.
Molecules na-emepụta akara eletrik dị iche iche dịka nhazi kemịkalụ si dị.
RA na-enweta nchọta usoro n'oge site na ịkpọ oku.
Ịrụ usoro transcriptome ogologo zuru oke
√ Saturation data
A chọrọ ịgụ 7 ugboro ole na ole iji ruo njupụta data atụnyere.
√ Identification Structure Transcript
Nchọpụta ụdị nhazi dị iche iche yana nkwenye zuru oke na-agụ nke ọ bụla.
√ Nlebanya ọkwa dị iche iche nke transcript - Gosipụta mgbanwe zoro ezo site na mkpirisi ọgụgụ
Ntụaka
Tang AD, Soulette CM, Baren MJV, et al.Nkọwa ederede zuru oke nke SF3B1 ngbanwe na leukemia lymphocytic na-adịghị ala ala na-ekpughe mbelata nke introns ejidere [J].Nkwukọrịta okike.
Teknụzụ na isi ihe bu n'obi kesaa ngwa kacha aga nke ọma na nso nso a nke teknụzụ usoro usoro dị elu dị iche iche na mpaghara nyocha dị iche iche yana echiche mara mma na nhazi nnwale na ngwuputa data.
Oge nzipu: Jan-08-2022