<mark id="bahkc"><acronym id="bahkc"></acronym></mark>

          国产精品久久久久久不卡_欧美激情精品久久_99久久免费精品国产72精品九九_精品熟人妻一区二区三区四区不卡_国产麻豆精品一区二区三区

          ?
          生物資訊

          基因組的飽和編輯

          作者:admin 來(lái)源:Nature 發(fā)布時(shí)間: 2014-09-05 08:32  瀏覽次數(shù):
          購(gòu)買(mǎi)進(jìn)口儀器、試劑和耗材——就在始于2001年的畢特博生物 m.kjhfd.cn

           Nature:基因組的飽和編輯

          基因組學(xué)領(lǐng)域,對(duì)能夠迅速、廉價(jià)確定突變的功能后果的方法有很大需求。

          這篇論文介紹了對(duì)基因組區(qū)域進(jìn)行飽和誘變(目的是產(chǎn)生所有可能的突變)、同時(shí)保持自然內(nèi)源性染色體環(huán)境的一個(gè)方法。

          該方法利用由CRISPR/Cas9 RNA引導(dǎo)的分裂和multiplex homology引導(dǎo)的修復(fù),其用途通過(guò)用所有可能的六聚體來(lái)替換一個(gè)六堿基對(duì)基因組區(qū)域和用所有可能的單一核苷酸變體來(lái)生成同一個(gè)完整的外顯子在BRCA1 的18號(hào)外顯子內(nèi)得到了演示。

          研究人員還對(duì)一個(gè)必要基因(即DBR1)的一個(gè)非常保守的編碼區(qū)域進(jìn)行了飽和編輯。該方法有望能夠促進(jìn)對(duì)順式調(diào)控元素和反式作用因子的高分辨率功能分析以及對(duì)由臨床測(cè)序工作所報(bào)告的具有不確定意義的變異體的解讀。

          原文摘要:

          Saturation editing of genomic regions by multiplex homology-directed repair

          Gregory M. Findlay, Evan A. Boyle, Ronald J. Hause, Jason C. Klein & Jay Shendure

           Saturation mutagenesis—coupled to an appropriate biological assay—represents a fundamental means of achieving a high-resolution understanding of regulatory and protein-coding nucleic acid sequences of interest. However, mutagenized sequences introduced in trans on episomes or via random or “safe-harbour” integration fail to capture the native context of the endogenous chromosomal locus. This shortcoming markedly limits the interpretability of the resulting measurements of mutational impact. Here, we couple CRISPR/Cas9 RNA-guided cleavage with multiplex homology-directed repair using a complex library of donor templates to demonstrate saturation editing of genomic regions. In exon 18 of BRCA1, we replac a six-base-pair (bp) genomic region with all possible hexamers, or the full exon with all possible single nucleotide variants (SNVs), and measure strong effects on transcript abundance attributable to nonsense-mediated decay and exonic splicing elements. We similarly perform saturation genome editing of a well-conserved coding region of an essential gene, DBR1, and measure relative effects on growth that correlate with functional impact. Measurement of the functional consequences of large numbers of mutations with saturation genome editing will potentially facilitate high-resolution functional dissection of both cis-regulatory elements and trans-acting factors, as well as the interpretation of variants of uncertain significance observed in clinical sequencing.

          購(gòu)買(mǎi)進(jìn)口儀器、試劑和耗材——就在始于2001年的畢特博生物 m.kjhfd.cn

          所有試劑均用于科研 ???? 北京畢特博生物技術(shù)有限責(zé)任公司 版權(quán)所有
          服務(wù)熱線:010-82015225/400-833-9299 | 傳真:010-62015131 | E-mail:info@bitebo.com
          京ICP備05028556號(hào)-1 ??京公網(wǎng)安備11010802008747??

          亚洲av无码乱码在线观看裸奔_欧美激情精品久久_99久久免费精品国产72精品九九_精品熟人妻一区二区三区四区不卡

          <mark id="bahkc"><acronym id="bahkc"></acronym></mark>