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相关概念视频

CRISPR01:59

CRISPR

48.6K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
48.6K
CRISPR and crRNAs02:53

CRISPR and crRNAs

16.3K
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
16.3K
Homologous Recombination02:31

Homologous Recombination

49.9K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
49.9K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

5.9K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
5.9K

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相关实验视频

Updated: May 15, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

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定向进化扩大了CRISPR-Cas12a的基因组编辑能力.

Enbo Ma, Kai Chen, Honglue Shi

    bioRxiv : the preprint server for biology
    |April 8, 2025
    PubMed
    概括

    研究人员设计了CRISPR-Cas12a以识别新的DNA序列,扩大了基因组编辑能力. 这一突破扩大了对以前无法实现的治疗和农业应用的遗传目标的访问范围.

    科学领域:

    • 分子生物学分子生物学
    • 生物技术是生物技术.
    • 基因组学就是基因组学.

    背景情况:

    • CRISPR-Cas12a是一种强大的基因组编辑工具.
    • 它的实用性受到一个狭窄的原始空间体相邻动机 (PAM) 识别 (5'-TTTV-3') 的限制,将准限制在基因组的1%左右.

    研究的目的:

    • 为了克服CRISPR-Cas12a.a的PAM限制.
    • 为更广泛的基因组向设计具有扩展PAM识别的变体.

    主要方法:

    • 使用细菌试验进行定向进化.
    • 理性工程的 *Lachnospiraceae 细菌* Cas12a (LbCas12a). 这是一个很好的方法.
    • 生物化学和基于细胞的测试来表征变异.

    主要成果:

    • 识别了具有扩展PAM识别的LbCas12a变体,包括非正规的动机.
    • 开发了Flex-Cas12a,它可以识别5'-NYHV-3'PAMs.
    • 扩展DNA识别到人类基因组的25%.

    结论:

    • 工程 Cas12a 变体显著提高了基因组向多功能性.

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    Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
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    Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

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  • Flex-Cas12a可以访问以前无法访问的基因组位置.
  • 为治疗和农业基因组工程提供了新的机会.