• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

利用小鼠乳腺类器官进行功能遗传缺失筛选和候选治疗靶点的验证。

Functional genetic dropout screens and validation of candidate therapeutic targets using mouse mammary tumoroids.

机构信息

Division of Molecular Pathology, the Netherlands Cancer Institute, 1066CX Amsterdam, the Netherlands.

Oncode Institute, 1066CX Amsterdam, the Netherlands.

出版信息

STAR Protoc. 2022 Jan 26;3(1):101132. doi: 10.1016/j.xpro.2022.101132. eCollection 2022 Mar 18.

DOI:10.1016/j.xpro.2022.101132
PMID:35146447
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8801381/
Abstract

We have previously developed 3D tumoroids derived from genetically engineered mouse models (GEMMs) of BRCA1/2-deficient breast cancer. Here, we describe how to genetically modify tumoroids, use them for functional genetic dropout screens, and carry out orthotopic transplantation of modified tumoroids. The purpose of this protocol is to screen for therapeutic targets and allow rapid and straightforward validation of the candidate targets. For complete details on the use and execution of this protocol, please refer to Paes Dias et al. (2021b).

摘要

我们之前已经开发了源自 BRCA1/2 缺陷型乳腺癌基因工程小鼠模型 (GEMM) 的 3D 类器官。在这里，我们将描述如何对类器官进行基因修饰，如何将其用于功能基因敲除筛选，以及如何进行修饰后的类器官的原位移植。本方案的目的是筛选治疗靶点，并允许候选靶点的快速和直接验证。有关该方案使用和执行的完整详细信息，请参见 Paes Dias 等人 (2021b)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/d6062edd1c96/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/be57d4c2112a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/8f2b9f88917b/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/8abc5ab463e4/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/9a14b8049fbf/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/d6062edd1c96/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/be57d4c2112a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/8f2b9f88917b/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/8abc5ab463e4/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/9a14b8049fbf/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a90e/8801381/d6062edd1c96/gr4.jpg

相似文献

1
Functional genetic dropout screens and validation of candidate therapeutic targets using mouse mammary tumoroids.利用小鼠乳腺类器官进行功能遗传缺失筛选和候选治疗靶点的验证。
STAR Protoc. 2022 Jan 26;3(1):101132. doi: 10.1016/j.xpro.2022.101132. eCollection 2022 Mar 18.
2
3D bioprinted mammary organoids and tumoroids in human mammary derived ECM hydrogels.3D 生物打印的人乳腺来源细胞外基质水凝胶中的乳腺类器官和肿瘤类器官。
Acta Biomater. 2019 Sep 1;95:201-213. doi: 10.1016/j.actbio.2019.06.017. Epub 2019 Jun 21.
3
Breast cancer PDxO cultures for drug discovery and functional precision oncology.用于药物发现和功能精准肿瘤学的乳腺癌 PDxO 培养物。
STAR Protoc. 2023 Sep 15;4(3):102402. doi: 10.1016/j.xpro.2023.102402. Epub 2023 Jul 3.
4
Establishment and long-term culture of mouse mammary stem cell organoids and breast tumor organoids.建立和长期培养小鼠乳腺干细胞类器官和乳腺肿瘤类器官。
STAR Protoc. 2021 May 29;2(2):100577. doi: 10.1016/j.xpro.2021.100577. eCollection 2021 Jun 18.
5
Comparative oncogenomics identifies combinations of driver genes and drug targets in BRCA1-mutated breast cancer.比较肿瘤基因组学鉴定了 BRCA1 突变型乳腺癌中驱动基因和药物靶点的组合。
Nat Commun. 2019 Jan 23;10(1):397. doi: 10.1038/s41467-019-08301-2.
6
Genome-scale chemogenomic CRISPR screens in human cells using the TKOv3 library.使用 TKOv3 文库在人类细胞中进行全基因组化学基因组 CRISPR 筛选。
STAR Protoc. 2021 Feb 5;2(1):100321. doi: 10.1016/j.xpro.2021.100321. eCollection 2021 Mar 19.
7
A systematic review on the culture methods and applications of 3D tumoroids for cancer research and personalized medicine.关于用于癌症研究和个性化医疗的3D肿瘤类器官培养方法及应用的系统综述。
Cell Oncol (Dordr). 2025 Feb;48(1):1-26. doi: 10.1007/s13402-024-00960-8. Epub 2024 May 28.
8
Image-guided radiotherapy in an orthotopic mouse model of rectal cancer.图像引导放疗在直肠癌原位模型小鼠中的应用。
STAR Protoc. 2022 Dec 16;3(4):101749. doi: 10.1016/j.xpro.2022.101749. Epub 2022 Oct 5.
9
Protocol for derivation of organoids and patient-derived orthotopic xenografts from glioma patient tumors.从胶质瘤患者肿瘤中衍生类器官和患者来源的原位异种移植物的方案。
STAR Protoc. 2021 May 12;2(2):100534. doi: 10.1016/j.xpro.2021.100534. eCollection 2021 Jun 18.
10
Generation of glioblastoma patient-derived organoids and mouse brain orthotopic xenografts for drug screening.用于药物筛选的胶质母细胞瘤患者来源的类器官和小鼠脑原位异种移植物的生成。
STAR Protoc. 2021 Feb 18;2(1):100345. doi: 10.1016/j.xpro.2021.100345. eCollection 2021 Mar 19.

引用本文的文献

1
Multi-omics analysis reveals distinct non-reversion mechanisms of PARPi resistance in BRCA1- versus BRCA2-deficient mammary tumors.多组学分析揭示了 BRCA1 缺陷型与 BRCA2 缺陷型乳腺肿瘤中 PARPi 耐药的不同非逆转机制。
Cell Rep. 2023 May 30;42(5):112538. doi: 10.1016/j.celrep.2023.112538. Epub 2023 May 19.

本文引用的文献

1
Loss of nuclear DNA ligase III reverts PARP inhibitor resistance in BRCA1/53BP1 double-deficient cells by exposing ssDNA gaps.核 DNA 连接酶 III 的缺失通过暴露单链 DNA 缺口使 BRCA1/53BP1 双缺陷细胞对 PARP 抑制剂耐药性逆转。
Mol Cell. 2021 Nov 18;81(22):4692-4708.e9. doi: 10.1016/j.molcel.2021.09.005. Epub 2021 Sep 22.
2
Understanding and overcoming resistance to PARP inhibitors in cancer therapy.理解和克服癌症治疗中对 PARP 抑制剂的耐药性。
Nat Rev Clin Oncol. 2021 Dec;18(12):773-791. doi: 10.1038/s41571-021-00532-x. Epub 2021 Jul 20.
3
Comparative oncogenomics identifies combinations of driver genes and drug targets in BRCA1-mutated breast cancer.
比较肿瘤基因组学鉴定了 BRCA1 突变型乳腺癌中驱动基因和药物靶点的组合。
Nat Commun. 2019 Jan 23;10(1):397. doi: 10.1038/s41467-019-08301-2.
4
BRCA-deficient mouse mammary tumor organoids to study cancer-drug resistance.BRCA 缺陷型小鼠乳腺肿瘤类器官用于研究癌症耐药性。
Nat Methods. 2018 Feb;15(2):134-140. doi: 10.1038/nmeth.4535. Epub 2017 Dec 11.
5
PTEN Loss in E-Cadherin-Deficient Mouse Mammary Epithelial Cells Rescues Apoptosis and Results in Development of Classical Invasive Lobular Carcinoma.E-钙黏蛋白缺陷型小鼠乳腺上皮细胞中PTEN缺失可挽救细胞凋亡并导致经典浸润性小叶癌的发生。
Cell Rep. 2016 Aug 23;16(8):2087-2101. doi: 10.1016/j.celrep.2016.07.059. Epub 2016 Aug 11.
6
REV7 counteracts DNA double-strand break resection and affects PARP inhibition.REV7可对抗DNA双链断裂切除并影响聚（ADP-核糖）聚合酶抑制作用。
Nature. 2015 May 28;521(7553):541-544. doi: 10.1038/nature14328. Epub 2015 Mar 23.
7
MAGeCK enables robust identification of essential genes from genome-scale CRISPR/Cas9 knockout screens.MAGeCK能够从全基因组规模的CRISPR/Cas9基因敲除筛选中可靠地鉴定必需基因。
Genome Biol. 2014;15(12):554. doi: 10.1186/s13059-014-0554-4.
8
The role of tumour-stromal interactions in modifying drug response: challenges and opportunities.肿瘤-基质相互作用在改变药物反应中的作用：挑战与机遇。
Nat Rev Drug Discov. 2013 Mar;12(3):217-28. doi: 10.1038/nrd3870.
9
Controlled gene expression in primary Lgr5 organoid cultures.在原代 Lgr5 类器官培养物中进行基因表达的控制。
Nat Methods. 2011 Dec 4;9(1):81-3. doi: 10.1038/nmeth.1802.
10
Somatic inactivation of E-cadherin and p53 in mice leads to metastatic lobular mammary carcinoma through induction of anoikis resistance and angiogenesis.小鼠中E-钙黏蛋白和p53的体细胞失活通过诱导失巢凋亡抗性和血管生成导致转移性小叶乳腺癌。
Cancer Cell. 2006 Nov;10(5):437-49. doi: 10.1016/j.ccr.2006.09.013.