iTracer在人脑类器官中的谱系记录。

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/978-1-0716-4310-5_5

Eugenio Gentile, Ashley Maynard, Zhisong He, Barbara Treutlein

{"title":"iTracer在人脑类器官中的谱系记录。","authors":"Eugenio Gentile, Ashley Maynard, Zhisong He, Barbara Treutlein","doi":"10.1007/978-1-0716-4310-5_5","DOIUrl":null,"url":null,"abstract":"Induced pluripotent stem cell (iPSC)-derived organoids provide models to study human organ development. Single-cell transcriptomics enables highly resolved descriptions of cell states within these systems; however, approaches are needed to directly determine the lineage relationship between cells. Here we provide a detailed protocol (Fig. 1) for the application of iTracer (He Z, Maynard A, Jain A, et al., Nat Methods 19:90-99, 2022), a recently published lineage recorder that combines reporter barcodes with inducible CRISPR-Cas9 scarring and is compatible with single-cell and spatial transcriptomics. iTracer is used to explore clonality and lineage dynamics during brain organoid development. More broadly, iTracer can be adapted to any iPSC-derived culture system to dissect lineage dynamics during normal or perturbed development.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2886 ","pages":"85-101"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Lineage Recording in Human Brain Organoids with iTracer.\",\"authors\":\"Eugenio Gentile, Ashley Maynard, Zhisong He, Barbara Treutlein\",\"doi\":\"10.1007/978-1-0716-4310-5_5\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Induced pluripotent stem cell (iPSC)-derived organoids provide models to study human organ development. Single-cell transcriptomics enables highly resolved descriptions of cell states within these systems; however, approaches are needed to directly determine the lineage relationship between cells. Here we provide a detailed protocol (Fig. 1) for the application of iTracer (He Z, Maynard A, Jain A, et al., Nat Methods 19:90-99, 2022), a recently published lineage recorder that combines reporter barcodes with inducible CRISPR-Cas9 scarring and is compatible with single-cell and spatial transcriptomics. iTracer is used to explore clonality and lineage dynamics during brain organoid development. More broadly, iTracer can be adapted to any iPSC-derived culture system to dissect lineage dynamics during normal or perturbed development.\",\"PeriodicalId\":18490,\"journal\":{\"name\":\"Methods in molecular biology\",\"volume\":\"2886 \",\"pages\":\"85-101\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2025-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Methods in molecular biology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1007/978-1-0716-4310-5_5\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in molecular biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/978-1-0716-4310-5_5","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}

引用次数: 0

摘要

诱导多能干细胞（iPSC）衍生的类器官为研究人体器官发育提供了模型。单细胞转录组学能够高度解析这些系统内的细胞状态；然而，需要直接确定细胞之间的谱系关系的方法。在这里，我们为iTracer的应用提供了详细的方案（图1）（He Z, Maynard a, Jain a, et ., Nat Methods 19:90- 99,2022）， iTracer是一种最近发表的谱系记录器，将报告基因条形码与可诱导的CRISPR-Cas9瘢痕结合起来，与单细胞和空间转录组学兼容。iTracer用于探索脑类器官发育过程中的克隆和谱系动力学。更广泛地说，iTracer可以适用于任何ipsc衍生的培养系统，以解剖正常或受干扰发育期间的谱系动态。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

Lineage Recording in Human Brain Organoids with iTracer.

Induced pluripotent stem cell (iPSC)-derived organoids provide models to study human organ development. Single-cell transcriptomics enables highly resolved descriptions of cell states within these systems; however, approaches are needed to directly determine the lineage relationship between cells. Here we provide a detailed protocol (Fig. 1) for the application of iTracer (He Z, Maynard A, Jain A, et al., Nat Methods 19:90-99, 2022), a recently published lineage recorder that combines reporter barcodes with inducible CRISPR-Cas9 scarring and is compatible with single-cell and spatial transcriptomics. iTracer is used to explore clonality and lineage dynamics during brain organoid development. More broadly, iTracer can be adapted to any iPSC-derived culture system to dissect lineage dynamics during normal or perturbed development.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.