Genetic Engineering of Human Embryonic Stem Cells for Precise Cell Fate Tracing During Human Lineage Development

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2018 Nov 20

PMID 30449321

Citations 10

Authors

Zhenyu Chen

Xudong Ren

Xiangjie Xu

Xiaojie Zhang

Yi Hui

Zhongliang Liu

Lei Shi

Yujiang Fang

Lin Ma

Yang Liu

Daniel Terheyden-Keighley

Ling Liu

Xiaoqing Zhang

Affiliations

Soon will be listed here.

Abstract

It is highly desirable to specify human developmental principles in an appropriate human model with advanced genetic tools. However, genetically engineering human cells with lineage-tracing systems has not been achieved. Here we introduce strategies to construct lineage-tracing systems in human embryonic stem cells (hESCs). The AAVS1 locus was suitable for the integration of the conditional reporter. The Cre-LoxP and Flp-FRT systems were highly sensitive, which may cause inaccurate lineage labeling in human cells. The recombination sensitivity and tracing fidelity could be finely tuned by modification of the LoxP recombination site. Moreover, tamoxifen-controllable Cre-LoxP and Flp-FRT systems showed compelling advantages in tightly tracing human lineages temporally. In proof-of-principle experiments, we traced human PAX6 neuroectoderm cells and revealed their full neural lineage differentiation potency both in vitro and in vivo. Devising and optimizing of lineage-tracing systems in hESCs will thus set up a solid foundation for human developmental studies.

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