Pseudouridine-mediated branch site recognition/selection during pre-mRNA splicing

前 mRNA 剪接过程中假尿苷介导的分支位点识别/选择

基本信息

  • 批准号:
    10026485
  • 负责人:
  • 金额:
    $ 36.14万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2020
  • 资助国家:
    美国
  • 起止时间:
    2020-08-10 至 2024-04-30
  • 项目状态:
    已结题

项目摘要

We propose to solve two long-standing problems in the field of pre-mRNA splicing: (1) how does the branch site recognition region (BSRR) of U2, which is a single unique sequence, recognize (via base-pairing) a variety of branch site sequences (BSS) in pre-mRNAs during splicing? And (2) how does the pseudouridine (Ψ), which is abundantly present in the U2 BSRR, contribute to this recognition? To address these questions, we recently developed a screening system, allowing us to screen a library of pre-mRNAs with randomized BSS sequences, under various genetic backgrounds where the numbers and combinations of Ψs in the U2 BSRR are manipulated and controlled. Our initial screen has generated exciting preliminary results, placing us in a unique position to solve the aforementioned long-standing problems. Three specific aims are proposed. 1. To expand the list of BSS and its 5' adjacent sequences recognized by different U2 variants We will build on our preliminary results and continue to screen the library (randomized BSS). Given that we have also shown in our preliminary experiments that a 6-nucleotide sequence 5'-adjacent to the BSS in pre- mRNA is also recognized by the U2 BSRR, we will screen additional libraries of pre-mRNAs with this 5'- adjacent sequence being randomized, under different U2 (differ in Ψ) backgrounds. In doing so, we expect to obtain a long list of BSS and its 5' adjacent sequences selected by different U2 variants with different numbers and combinations of Ψs in the BSRR, thus helping decode how Ψs contribute to BSS recognition. 2. To evaluate whether/how Ψs within the U2 BSRR contribute to BSS selection / gene expression With the BSS and its 5' adjacent sequences handy, we will search for the selected sequences in naturally occurring pre-mRNAs. The splicing efficiency of these pre-mRNAs will be tested under U2 variants (differ in Ψ). In doing so, we can link Ψ in the U2 BSRR to splicing/gene regulation. We also expect that some U2 variants prefer one BSS+5' adjacent sequence over another, whereas other U2 variants have completely opposite preference between the same two BSS+5' adjacent sequences. We will create several constructs with two BSS+5' adjacent sequences being placed in parallel, and test the usage of one BSS over the other during splicing under specific U2 backgrounds, thus linking Ψ-mediated BSS selection to alternative splicing. 3. To dissect how individual U2 RNA variants recognize various BSSs and their 5'-adjacent sequences Due to incomplete pseudouridylation at any given site, there exists a mixture of U2 variants (differ in Ψ) in cells. We will dissect, in detail, how the mixture of U2 variants individually recognizes a specific BSS during splicing. We will use the Oxford nanopore technology to quantitatively map, at single molecule level, Ψ in the BSRR of U2 isolated from the spliceosomes, which are assembled onto a pre-mRNA with a specific BSS+5' adjacent sequence. In doing so, we can assess how each individual U2 variant behaves, in a natural context, in the process of BSS recognition. A complete picture of Ψ-mediated BSS recognition is expected to emerge.
我们提出要解决两个长期存在的问题,在该领域的前mRNA剪接:(1)如何分支 U2的位点识别区(BSRR)是一个单一的独特序列,识别(通过碱基配对) 剪接过程中前体mRNA中的各种分支位点序列(BSS)?以及(2)假尿苷如何 (1),这是大量存在于U2 BSRR,有助于这种认识?为了解决这些问题, 我们最近开发了一个筛选系统,允许我们用随机化的 BSS序列,在不同的遗传背景下,其中U2中的碱基的数量和组合 BSRR被操纵和控制。我们的初步筛选产生了令人兴奋的初步结果,使我们 在解决上述长期存在的问题方面处于独特地位。提出了三个具体目标。 1.为了扩展由不同U2变体识别的BSS及其5'相邻序列的列表, 我们将在初步结果的基础上继续筛选文库(随机BSS)。鉴于我们 在我们的初步实验中也表明,在前- mRNA也被U2 BSRR识别,我们将用这种5 '- 相邻序列被随机化,在不同的U2(不同的U2)背景下。在这样做时,我们希望 获得BSS及其5'相邻序列的长列表,所述BSS及其5'相邻序列由具有不同 BSRR中Ψ的数量和组合,从而帮助解码Ψ如何对BSS识别做出贡献。 2.评价U2 BSRR内的BSs是否/如何促进BSS选择/基因表达 有了BSS及其5'端相邻序列,我们将自然地搜索所选序列。 前体mRNA将在U2变体下测试这些前mRNA的剪接效率(不同之处在于 )。通过这样做,我们可以将U2 BSRR中的Ψ与剪接/基因调节联系起来。我们还预计一些U2 变体偏好一个BSS+5'相邻序列而不是另一个,而其他U2变体具有完全的 相同的两个BSS+5'相邻序列之间的相反偏好。我们将创建几个结构 两个BSS+5'相邻序列并行放置,并测试一个BSS相对于另一个BSS的使用情况 在特定的U2背景下的剪接过程中,从而将β-介导的BSS选择与选择性剪接联系起来。 3.剖析单个U2 RNA变体如何识别各种BSS及其5 '-邻近序列 由于在任何给定位点处的不完全假尿苷化,存在U2变体的混合物(在碱基上不同), 细胞我们将详细分析U2变体的混合物如何在特定的BSS中单独识别特定的BSS。 拼接我们将使用牛津纳米孔技术,在单分子水平上定量绘制, 从剪接体分离的U2的BSRR,剪接体组装到具有特异性BSS+5'端的前体mRNA上。 相邻序列在这样做的过程中,我们可以评估每个U2变体在自然环境中的行为, 在BSS识别的过程中。一个完整的图片,预计将出现的BSS介导的识别。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

YI-TAO YU其他文献

YI-TAO YU的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('YI-TAO YU', 18)}}的其他基金

Pseudouridine-mediated branch site recognition/selection during pre-mRNA splicing
前 mRNA 剪接过程中假尿苷介导的分支位点识别/选择
  • 批准号:
    10383755
  • 财政年份:
    2020
  • 资助金额:
    $ 36.14万
  • 项目类别:
Pseudouridine-mediated branch site recognition/selection during pre-mRNA splicing
前 mRNA 剪接过程中假尿苷介导的分支位点识别/选择
  • 批准号:
    10612829
  • 财政年份:
    2020
  • 资助金额:
    $ 36.14万
  • 项目类别:
Pseudouridine-mediated branch site recognition/selection during pre-mRNA splicing
前 mRNA 剪接过程中假尿苷介导的分支位点识别/选择
  • 批准号:
    10231213
  • 财政年份:
    2020
  • 资助金额:
    $ 36.14万
  • 项目类别:
Suppression of disease causingnonsense mutations by targeted mRNA pseudouridylation
通过靶向 mRNA 假尿苷化抑制引起无义突变的疾病
  • 批准号:
    9805188
  • 财政年份:
    2019
  • 资助金额:
    $ 36.14万
  • 项目类别:
Expanding the genetic code by targeted pseudouridylation
通过靶向假尿苷化扩展遗传密码
  • 批准号:
    8550118
  • 财政年份:
    2012
  • 资助金额:
    $ 36.14万
  • 项目类别:
Expanding the genetic code by targeted pseudouridylation
通过靶向假尿苷化扩展遗传密码
  • 批准号:
    8419298
  • 财政年份:
    2012
  • 资助金额:
    $ 36.14万
  • 项目类别:
Regulation of telomerase activity and aging by 2'-O-methylation in telomerase RNA
端粒酶RNA中2-O-甲基化对端粒酶活性和衰老的调节
  • 批准号:
    8516934
  • 财政年份:
    2012
  • 资助金额:
    $ 36.14万
  • 项目类别:
Expanding the genetic code by targeted pseudouridylation
通过靶向假尿苷化扩展遗传密码
  • 批准号:
    8914641
  • 财政年份:
    2012
  • 资助金额:
    $ 36.14万
  • 项目类别:
Expanding the genetic code by targeted pseudouridylation
通过靶向假尿苷化扩展遗传密码
  • 批准号:
    8720024
  • 财政年份:
    2012
  • 资助金额:
    $ 36.14万
  • 项目类别:
Regulation of telomerase activity and aging by 2'-O-methylation in telomerase RNA
端粒酶RNA中2-O-甲基化对端粒酶活性和衰老的调节
  • 批准号:
    8242209
  • 财政年份:
    2012
  • 资助金额:
    $ 36.14万
  • 项目类别:

相似海外基金

How Does Particle Material Properties Insoluble and Partially Soluble Affect Sensory Perception Of Fat based Products
不溶性和部分可溶的颗粒材料特性如何影响脂肪基产品的感官知觉
  • 批准号:
    BB/Z514391/1
  • 财政年份:
    2024
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Training Grant
BRC-BIO: Establishing Astrangia poculata as a study system to understand how multi-partner symbiotic interactions affect pathogen response in cnidarians
BRC-BIO:建立 Astrangia poculata 作为研究系统,以了解多伙伴共生相互作用如何影响刺胞动物的病原体反应
  • 批准号:
    2312555
  • 财政年份:
    2024
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Standard Grant
RII Track-4:NSF: From the Ground Up to the Air Above Coastal Dunes: How Groundwater and Evaporation Affect the Mechanism of Wind Erosion
RII Track-4:NSF:从地面到沿海沙丘上方的空气:地下水和蒸发如何影响风蚀机制
  • 批准号:
    2327346
  • 财政年份:
    2024
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Standard Grant
Graduating in Austerity: Do Welfare Cuts Affect the Career Path of University Students?
紧缩毕业:福利削减会影响大学生的职业道路吗?
  • 批准号:
    ES/Z502595/1
  • 财政年份:
    2024
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Fellowship
感性個人差指標 Affect-X の構築とビスポークAIサービスの基盤確立
建立个人敏感度指数 Affect-X 并为定制人工智能服务奠定基础
  • 批准号:
    23K24936
  • 财政年份:
    2024
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Insecure lives and the policy disconnect: How multiple insecurities affect Levelling Up and what joined-up policy can do to help
不安全的生活和政策脱节:多种不安全因素如何影响升级以及联合政策可以提供哪些帮助
  • 批准号:
    ES/Z000149/1
  • 财政年份:
    2024
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Research Grant
How does metal binding affect the function of proteins targeted by a devastating pathogen of cereal crops?
金属结合如何影响谷类作物毁灭性病原体靶向的蛋白质的功能?
  • 批准号:
    2901648
  • 财政年份:
    2024
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Studentship
Investigating how double-negative T cells affect anti-leukemic and GvHD-inducing activities of conventional T cells
研究双阴性 T 细胞如何影响传统 T 细胞的抗白血病和 GvHD 诱导活性
  • 批准号:
    488039
  • 财政年份:
    2023
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Operating Grants
New Tendencies of French Film Theory: Representation, Body, Affect
法国电影理论新动向:再现、身体、情感
  • 批准号:
    23K00129
  • 财政年份:
    2023
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
The Protruding Void: Mystical Affect in Samuel Beckett's Prose
突出的虚空:塞缪尔·贝克特散文中的神秘影响
  • 批准号:
    2883985
  • 财政年份:
    2023
  • 资助金额:
    $ 36.14万
  • 项目类别:
    Studentship
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了