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Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

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...

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Updated: Jun 21, 2025

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

工程微型 IscB 尼卡斯,用于强大的基础编辑,具有广泛的准范围.

Linxiao Han¹, Yueer Hu¹, Qiqin Mo¹

¹Gene Editing Center, School of Life Science and Technology, ShanghaiTech University, Shanghai, China.

Nature chemical biology

|July 8, 2024

概括

研究人员从IscB设计了微型基因编辑器IminiBEs,以实现高效的基因组编辑. 融合Sso7d蛋白质创造了SIminiBEs,进一步提高了编辑效率,并扩大了对精确基因修改的目标识别.

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Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

相关实验视频

Last Updated: Jun 21, 2025

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物技术是生物技术.

背景情况:

IscB是一种Cas9同类物,具有较小的尺寸,有利于病毒传递系统.
本地OgeuIscB在哺乳动物细胞中表现出低的编辑效率,限制了其治疗潜力.

研究的目的:

来自OgeuIscB的高效微型基因编辑器,以提高基因组突变能力.
为了提高编辑效率和基于IscB的基础编辑器的目标站点识别.

主要方法:

工程 OgeuIscB 尼克酶及其同源 ωRNA 创建 IminiBEs (微型基础编辑器).
将Sso7dDNA结合蛋白与IminiBEs的融合产生SIminiBEs.
评估哺乳动物细胞中IminiBEs和SIminiBEs的编辑效率和目标相邻的图案识别.

主要成果:

IMiniCBE的平均编辑效率为67%,IminiABE的平均效率为52%.
在具有挑战性的网站上,SIminiBEs的编辑效率提高了两到三倍.
IMiniCBE和SIminiCBE扩大了目标识别的范围,超出了正规的NWRRNA动机,包括NNRR,NNRY和NNYR.

结论:

IMiniBEs和SIminiBEs代表了高效和多功能迷你基础编辑工具.
这些工程编辑器促进了特定站点的基因组突变,提高了效率和更广泛的准范围.
这项开发对先进的基因编辑应用有前途,需要精确的基因组改变.