Tracing Cell Lineages

描述(申请人提供)：成人人体由数万亿个细胞组成，这些细胞都来自一个受精的卵细胞。在成人中，大多数组织处于一种 不断的流动，旧细胞死亡，新细胞从常驻的干细胞群体中产生，以神秘的方式改变它们的身份。细胞群体变化的流动性从我们受孕的那一刻开始--直到我们生命的最后一口气。多细胞生命在一个高度有序的过程的音乐中起舞，指挥的乐谱并不为人所知。当细胞失去方向，并以不受控制的方式分裂，失去身份时，就会出现癌症等疾病。其他疾病的特点是细胞丢失，由细胞凋亡或自身免疫等不必要的自我消除引发。生物学中最大的斗争之一是在细胞和分子水平上理解这些基本过程。我们的想法是开发一种新的工具，利用一种新的DNA条形码技术，帮助我们追踪单个细胞的祖先。这项提议描述了一种追踪个体细胞进化历史的新方法--在最可能的颗粒水平上，个体细胞。我们利用新技术(深度测序和可编程DNA结合酶)，将它们结合在一起，创建单个细胞谱系示踪器，每个细胞在其中写入自己唯一的条形码。这个系统由一台分子打字机组成，每次细胞进入细胞周期时，打字机都会输入唯一的条形码。此外，这种书写的条形码会随着未来的每一代人而积累。我们的愿景是将该系统引入受精受精卵中，以培育出小鼠品系。从本质上讲，老鼠体内的每个细胞都会包含一个独特的条形码，每个条形码都会包含有关其祖先血统的信息，可以追溯到最初受精的受精卵。我们相信，开发一种定义单细胞遗传谱系的量化方法将改变下一代发育和疾病生物学家处理问题的方式。

DESCRIPTION (provided by applicant): The adult human body is composed of trillions of cells that all originated from a single fertilized egg cell. In the adult, most tissues are in a state of constant flux, where old cells die and new cells are created from resident populations of stem cells, changing their identities in mysterious ways. The fluidity of change among cell populations initiate from the moment we are conceived- to our final breath of life. Multicellular life dances t the music of a highly ordered process, directed by a score that is not well understood. Disease such as cancer emerges when cells lose their directions, and divide in an uncontrolled manner, losing their identities. Other diseases are hallmarked by a loss of cells, triggered by unwanted self-elimination such as apoptosis or autoimmunity. One of the greatest struggles in biology is to understand these fundamental processes, at the cellular and molecular level. Our idea is to develop a new tool that will help us track the ancestry of individual cells, using a novel DNA barcoding technology. This proposal describes a new approach for tracking the evolutionary history of individual cells- at the most possible granular level, the individual cells. We take advantage of new technologies (deep sequencing and programmable DNA binding enzymes), combining them in a way to create a single cell lineage tracer in which each cell writes its own unique barcode. This system is comprised of a molecular 'typewriter' that types a unique barcode every time a cell goes into cell cycle. Moreover, this written barcode accumulates with each future generation. Our vision is to introduce this system into fertlized zygotes, were mouse lines will be developed. In essence every cell in the mouse will contain a unique barcode, and each barcode would contain information on its ancestral lineage, tracing back to the initial fertilized zygote. We believe that developing a quantitative approach for defining single cell inheritance lineages will transform how the next generation of developmental and disease biologists approach their problems.