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相关概念视频

Cloning of Dolly the Sheep01:08

Cloning of Dolly the Sheep

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The first successfully cloned mammal was Dolly, a sheep, born on 5th July 1996 at Roslin Institute, Scotland. The cloned sheep was named after the American singer Dolly Parton. Dolly lived for seven years and died of respiratory complications, which is speculated to be due to the actual age of her DNA. Because the DNA in cloned cells belongs to an older individual,  the cloned individual’s life expectancy may be affected. Indeed, analysis of Dolly’s DNA revealed shorter...
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Reproductive Cloning01:27

Reproductive Cloning

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Reproductive cloning is the process of producing a genetically identical copy—a clone—of an entire organism. While clones can be produced by splitting an early embryo—similar to what happens naturally with identical twins—cloning of adult animals is usually done by a process called somatic cell nuclear transfer (SCNT).
Somatic Cell Nuclear Transfer
In SCNT, an egg cell is taken from an animal and its nucleus is removed, creating an enucleated egg. Then a somatic...
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Long-patch Base Excision Repair01:02

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Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:
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Introduction to Nuclear Reprogramming01:14

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Nuclear reprogramming is the process of switching gene expression of one cell type to that of another cell type, usually from a differentiated cell state to an undifferentiated cell state. Differentiation occurs during processes such as development and morphogenesis, tissue regeneration, and malignancy. Cells can also be artificially induced to reprogram their gene expression by techniques such as nuclear transfer, induced pluripotency, and cell fusion. Such techniques have many applications in...
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Genome Annotation and Assembly03:36

Genome Annotation and Assembly

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The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.
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Improving Student Outcomes with an Adaptable Molecular Cloning Course-Based Undergraduate Research Experience
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改进了TALE组装的用户克隆,并将其应用于基础编辑.

Jizeng Zhou1, Jiaowei Wang2, Fangbing Chen2

  • 1School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou, China.

PloS one
|August 4, 2023
PubMed
概括

一种新的USER克隆方法简化了转录激活器样效应器 (TALEs) 的构建. 这种快速的技术可以实现高效的基准编辑,减少非目标DNA效应,有利于实验室应用.

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科学领域:

  • 分子生物学分子生物学
  • 基因编辑技术的技术
  • 合成生物学 合成生物学

背景情况:

  • 转录激活器样效应器 (TALEs) 是基因组编辑,转录调节和DNA成像的多功能工具.
  • 传统的TALE构造复杂且耗时,限制了它们在实验室的常规使用.

研究的目的:

  • 开发一种简单,快速和实验室友好的TALE组装方法.
  • 使用改进的TALE构造技术创建高效的基础编辑系统.

主要方法:

  • 使用乌拉特异性切除试剂 (USER) 克隆用于TALE组装.
  • 使用TALE剪切模板和含有脱氧氨的原料的聚合酶连锁反应.
  • 在层次上重新加入了使用USER和T4DNA结合酶进行高效向量构造的TALE剪切单元.

主要成果:

  • 通过使用USER组装方法成功构建了TALE-脱氨酶.
  • 证明了高效的C-to-T和A-to-G基编辑,当TALE-deaminases与Cas9尼克酶相结合时.
  • 在基准编辑应用中消除了可预测的DNA脱效应.

结论:

  • 改进的USER组件是TALE构建的高效和快速技术.
  • 这种方法简化了基于TALE的基因组工程和基因编辑.
  • 该技术为DNA向应用提供了一种有价值的,实验室友好的方法.