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基于 PiggyBac 的隐性筛选方法鉴定多能性调控因子。

A PiggyBac-based recessive screening method to identify pluripotency regulators.

机构信息

Wellcome Trust Centre for Stem Cell Research, Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.

出版信息

PLoS One. 2011 Apr 18;6(4):e18189. doi: 10.1371/journal.pone.0018189.

DOI:10.1371/journal.pone.0018189
PMID:21533166
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3078921/
Abstract

Phenotype driven genetic screens allow unbiased exploration of the genome to discover new biological regulators. Bloom syndrome gene (Blm) deficient embryonic stem (ES) cells provide an opportunity for recessive screening due to frequent loss of heterozygosity. We describe a strategy for isolating regulators of mammalian pluripotency based on conversion to homozygosity of PiggyBac gene trap insertions combined with stringent selection for differentiation resistance. From a screen of 2000 mutants we obtained a disruptive integration in the Tcf3 gene. Homozygous Tcf3 mutants showed impaired differentiation and enhanced self-renewal. This phenotype was reverted in a dosage sensitive manner by excision of one or both copies of the gene trap. These results provide new evidence confirming that Tcf3 is a potent negative regulator of pluripotency and validate a forward screening methodology to identify modulators of pluripotent stem cell biology.

摘要

表型驱动的基因筛选允许对基因组进行无偏探索，以发现新的生物调节剂。布卢姆综合征基因 (Blm) 缺陷型胚胎干细胞 (ES) 细胞由于经常发生杂合性丢失，提供了隐性筛选的机会。我们描述了一种基于转座子基因捕获插入物纯合化的策略，结合严格选择分化抗性，以分离哺乳动物多能性的调节剂。在对 2000 个突变体进行筛选后，我们在 Tcf3 基因中获得了一个破坏性的整合。Tcf3 纯合子突变体显示出分化受损和自我更新增强。通过基因捕获的一个或两个拷贝的缺失，可以以剂量敏感的方式逆转这种表型。这些结果提供了新的证据，证实 Tcf3 是多能性的一个强有力的负调节剂，并验证了一种正向筛选方法来鉴定多能干细胞生物学的调节剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/04be3ddde994/pone.0018189.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/d2cf61e3870f/pone.0018189.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/e69d09edf14e/pone.0018189.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/454bfbd231a6/pone.0018189.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/858585665b36/pone.0018189.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/cec84ad0a9b3/pone.0018189.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/a77aa61a6601/pone.0018189.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/04be3ddde994/pone.0018189.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/d2cf61e3870f/pone.0018189.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/e69d09edf14e/pone.0018189.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/454bfbd231a6/pone.0018189.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/858585665b36/pone.0018189.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/cec84ad0a9b3/pone.0018189.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/a77aa61a6601/pone.0018189.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3b9c/3078921/04be3ddde994/pone.0018189.g007.jpg

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Nat Cell Biol. 2011 Jun 19;13(7):838-45. doi: 10.1038/ncb2267.
2
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3
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EMBO J. 2020 Jan 15;39(2):e102591. doi: 10.15252/embj.2019102591. Epub 2019 Nov 29.
4
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Cell Stem Cell. 2019 May 2;24(5):785-801.e7. doi: 10.1016/j.stem.2019.03.017. Epub 2019 Apr 25.
5
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6
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8
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