YAN ADAM
dabi'ar halitta
Tsarin dogon karantawa yana gano GGC maimaita faɗaɗawa a cikin NOTCH2NLC da ke da alaƙa da cutar haɗar ƙwayar cuta ta neuronal.
Maimaita ONT |Illumina |Gabaɗaya exome jeri |CRISPR-Cas9 ONT jerin niyya |RNA-seq |ONT 5mC methylation kira
Karin bayanai
1.Ta hanyar bincike na Linkage akan babban dangin NIID, an gano yankuna biyu masu alaƙa.
2.ONT-based dogon karanta jerin abubuwan da aka yi da Cas-9 mai daidaitawa ONT jerin abubuwan haɓakawa sun gano yuwuwar sanadin kwayoyin halitta na NIID, GGC maimaita faɗaɗa a cikin 5′ UTR na NOTCH2NLC.Wannan binciken ya ba da rahoton maimaita faɗaɗawa a cikin takamaiman ƙwayoyin halittar ɗan adam a karon farko waɗanda suka samo asali ta hanyar kwafi.
3.RNA jerin abubuwan da aka saukar sun bayyana abubuwan da ba a saba gani ba a farkon ko cikin GGC maimaita fadada yankuna a cikin NOTCH2NLC.
Fage
NCutar haɗar ƙwayar cuta ta Euronal (NIID) cuta ce mai ci gaba kuma mai saurin mutuwa, wacce ke da alaƙa da kasancewar eosinophilic hyaline intranuclear inclusions a cikin tsarin tsakiya da na gefe.Bayyanuwansa masu saurin canzawa suna haifar da matsaloli masu yawa a cikin ganewar asali har sai an gabatar da kwayar cutar fata.Duk da haka, hanyoyin da suka samo asali na histopathology har yanzu suna fama da rashin ganewa, wanda ke kira ga fahimtar kwayoyin halitta na NIID.
Nasarorin da aka samu
Binciken haɗin gwiwa
SHort-read sequencing based whole genome sequencing (WGS) da cikakken exome sequencing (WES) an yi shi akan babban dangin NIID (an shafa 13 da mambobi 7 da basu shafa ba).Binciken haɗin kai akan SNPs da aka fitar daga waɗannan bayanan sun bayyana yankuna biyu kawai masu alaƙa: yanki na 3.5 Mb a 1p36.31-p36.22 (mafi girman LOD = 2.32) da yankin 58.1 Mb a 1p22.1-q21.3 (mafi girman LOD: 4.21). ).Koyaya, ba a gano SNPs ko CNVs masu cutarwa ba a cikin waɗannan yankuna masu alaƙa.
GGC maimaita fadadawa a cikin NOTCH2NLC
NAn gudanar da jerin abubuwan da aka yi amfani da su a kan 13 da abin ya shafa da kuma 4 da ba a shafa ba daga iyalai 8 (wani memba da abin ya shafa an tsara shi ta hanyar dandalin Pacbio na dogon lokaci.).Bayanan da aka dade ana karantawa sun bayyana cutar da ke da alaƙa da GGC maimaituwa a cikin 5′ UTR na taswirar halittar NOTCH2NLC zuwa yanki mai haɗin 58.1 Mb (Hoto 1).Hakanan an gano waɗannan maimaitawa a cikin duk lokuta 40 na NIID na lokaci-lokaci da RP-PCR suka gwada.
Cas-9 matsakaicin matsakaicin manufa akan dandamali na nanopore an yi aiki don cimma babban ɗaukar hoto akan maimaita NOTCH2NLC (100 X-1,795 X).Waɗannan jerin ra'ayoyin sun yarda da kyau tare da binciken da aka yi a baya akan maimaitawar GGC.Bugu da ƙari, an gano maimaitawar {(GGA) n (GGC) n} a matsayin alama mai yuwuwar alamar halitta don rashin ƙarfi-mafi rinjayen phenotype (Hoto 2).
Hoto 1. Cutar da ke hade da sake faɗaɗa da aka gano akan exon 1 na NOTCH2NLC isoforms.
Hoto 2. Matsalolin yarjejeniya na NPTCH2NLC maimaitawa a cikin marasa lafiya na NIID tare da (*) ko kuma ba tare da rauni-mafi yawan phenotype ba.
NKwayoyin OTCH2NL kwayoyin halitta ne na mutum, waɗanda aka yi imanin suna taka muhimmiyar rawa a cikin juyin halittar kwakwalwar ɗan adam da cututtukan jijiya.Duk da haka, kwayoyin halitta NOTCH2 guda uku (NOTCH2NLA, NOTCH2NLB da NOTCH2NLC) tare da> 99.1% jerin sunayen ba a warware su ba har sai sabon tsarin halittar ɗan adam.Ba tare da haɗin gwiwa ba da dogon karanta jerin abubuwan da ke kan dandamali na nanopore sun nuna fa'idodi masu ban sha'awa a cikin warware yankuna masu kamanni da (GGC) n maimaita tare da 100% GC-arziƙi.
GGC maimaita fadadawa a cikin NOTCH2NLC
TAn aiwatar da jerin abubuwan da aka rubuta akan mambobi 2 da abin ya shafa da 2 da ba a shafa ba.An ƙididdige zurfin karantawa daidai gwargwado akan ma'ana da madaidaicin madauri a sama na farkon exons na NOTCH2NL paralogs.An samo rubutun anti-hankali mara kyau kawai a cikin lamuran da abin ya shafa, waɗanda ke zaune a farkon ko a cikin yanki mai maimaita (Purple kololuwa a F1-14 da F1-16 a cikin Hoto 3.).Bugu da ƙari, an gano 54 DEGs kuma duk an wadatar da su a cikin GO da MPO sharuddan da suka shafi ayyukan neuronal.
Hoto 3. Daidaitaccen zurfin karantawa akan sama na farkon exon na NOTCH2NLC a cikin abubuwan da ba su da tasiri (a sama) da abin da aka shafa (a ƙasa).
Fasaha
Oxford Nanopore Teghnologies (ONT)
Nsequencing na anopore yana bambanta kansa da sauran dandamali na jerin abubuwa, domin ana karanta nucleotide kai tsaye ba tare da tsarin haɗin DNA ba.Yayin da DNA guda ɗaya ke wucewa ta cikin nau'in furotin mai girman nano (nanopore), nucleotides daban-daban suna haifar da igiyoyin ionic daban-daban, waɗanda za'a iya kama su kuma a tura su cikin jerin tushe.ONT tsarin dandamali kanta baya nuna ƙayyadaddun ƙayyadaddun fasaha akan tsawon karatun DNA.Saboda haka, Ultra-long reads (ULRs) suna samuwa don haɗuwar genome mai inganci.Haka kuma, waɗannan dogon karatu masu tsayi, waɗanda ke da tsayin daka don ketare sifofi masu sarƙaƙƙiya ko bambance-bambancen tsari, suna taimakawa wajen shawo kan iyakokin taƙaitaccen karatun anan.
Tsarin Nanopore
Bambancin tsarin (SV).
Sjerin abubuwan da ba su da haɗin kai an adana bayanan methylation na DNA akan samfuri.Methylated A, T, C da G suna haifar da igiyoyin ionic daban-daban daga waɗanda ba su da methylated, waɗanda za a iya karanta su kai tsaye ta dandamali.Mabiyan Nanopore yana ba da ikon yin bayanin gaba ɗaya-genome na 5mC da 6mA a ƙudurin nucleotide guda ɗaya.
Magana
Jun Sone, et.al.Tsawon dogon karantawa yana gano GGC maimaita faɗaɗa a cikin NOTCH2NLC da ke da alaƙa da cututtukan haɗar ƙwayoyin cuta na neuronal.Halitta Halitta (2019)
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-06-2022