An in vivo CRISPR screen in chick embryos reveals a role for MLLT3 in specification of neural cells from the caudal epiblast.

IF 3.7 2区生物学 Q1 DEVELOPMENTAL BIOLOGY

Development Pub Date : 2025-02-01 Epub Date: 2025-02-12 DOI:10.1242/dev.204591

Ashley R G Libby, Tiago Rito, Arthur Radley, James Briscoe

引用次数: 0

Abstract

Tissue development relies on the coordinated differentiation of stem cells in dynamically changing environments. The formation of the vertebrate neural tube from stem cells in the caudal lateral epiblast is a well-characterized example. Despite an understanding of the signalling pathways involved, the gene regulatory mechanisms remain poorly defined. To address this, we developed a multiplexed in vivo CRISPR screening approach in chick embryos targeting genes expressed in the caudal epiblast and neural tube. This revealed a role for MLLT3, a component of the super elongation complex, in the specification of neural fate. Perturbation of MLLT3 disrupted neural tube morphology and reduced neural fate acquisition. Mutant forms of retinoic acid receptor A lacking the MLLT3 binding domain similarly reduced neural fate acquisition. Together, these findings validate an in vivo CRISPR screen strategy in chick embryos and identify a previously unreported role for MLLT3 in caudal neural tissue specification.

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鸡胚胎的体内CRISPR筛选揭示了MLLT3在尾端外胚层神经细胞分化中的作用。

组织发育依赖于干细胞在动态变化环境中的协调分化。脊椎动物神经管是由尾侧上胚层（CLE）的干细胞形成的，这是一个特征鲜明的例子。尽管我们已经了解了其中的信号通路，但基因调控机制仍不甚明了。为了解决这个问题，我们在小鸡胚胎中开发了一种针对在尾侧上胚层和神经管中表达的基因的多重体内 CRISPR 筛选方法。这揭示了超级伸长复合体的一个组成部分 MLLT3 在神经命运的规范中的作用。干扰 MLLT3 会破坏神经管形态并减少神经命运的获得。缺乏 MLLT3 结合域的视黄酸受体 A 突变体也同样减少了神经命运的获得。这些发现共同验证了在小鸡胚胎中进行体内 CRISPR 筛选的策略，并确定了 MLLT3 在尾部神经组织规格化中以前未报道的作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Development 生物-发育生物学

CiteScore

6.70

自引率

4.30%

发文量

433

审稿时长

3 months

期刊介绍： Development’s scope covers all aspects of plant and animal development, including stem cell biology and regeneration. The single most important criterion for acceptance in Development is scientific excellence. Research papers (articles and reports) should therefore pose and test a significant hypothesis or address a significant question, and should provide novel perspectives that advance our understanding of development. We also encourage submission of papers that use computational methods or mathematical models to obtain significant new insights into developmental biology topics. Manuscripts that are descriptive in nature will be considered only when they lay important groundwork for a field and/or provide novel resources for understanding developmental processes of broad interest to the community. Development includes a Techniques and Resources section for the publication of new methods, datasets, and other types of resources. Papers describing new techniques should include a proof-of-principle demonstration that the technique is valuable to the developmental biology community; they need not include in-depth follow-up analysis. The technique must be described in sufficient detail to be easily replicated by other investigators. Development will also consider protocol-type papers of exceptional interest to the community. We welcome submission of Resource papers, for example those reporting new databases, systems-level datasets, or genetic resources of major value to the developmental biology community. For all papers, the data or resource described must be made available to the community with minimal restrictions upon publication. To aid navigability, Development has dedicated sections of the journal to stem cells & regeneration and to human development. The criteria for acceptance into these sections is identical to those outlined above. Authors and editors are encouraged to nominate appropriate manuscripts for inclusion in one of these sections.