Marker-free genetic manipulations in yeast using CRISPR/CAS9 system

被引：15

作者：

Soreanu, Inga ^{[1
,2
]}

Hendler, Adi ^{[1
,2
]}

Dahan, Danielle ^{[1
,2
]}

Dovrat, Daniel ^{[1
,2
]}

Aharoni, Amir ^{[1
,2
]}

机构：

[1] Ben Gurion Univ Negev, Dept Life Sci, IL-84105 Beer Sheva, Israel

[2] Ben Gurion Univ Negev, Natl Inst Biotechnol Negev, IL-84105 Beer Sheva, Israel

来源：

CURRENT GENETICS | 2018年 / 64卷 / 05期

基金：

欧盟地平线“2020”; 美国国家科学基金会;

关键词：

S; cerevisiae; CRISPR; CAS9; Yeast; Marker-free; SACCHAROMYCES-CEREVISIAE; OXIDATIVE STRESS; OFF-TARGET; GENOME; DISRUPTION; INTEGRATION; RESISTANCE; DELETION; CLONING; TRANSCRIPTION;

D O I：

10.1007/s00294-018-0831-y

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The budding yeast is currently one of the major model organisms for the study of a wide variety of biological processes. Genetic manipulation of yeast involves the extensive usage of selectable markers that can lead to undesired effects. Thus, marker-free genetic manipulation in yeast is highly desirable for gene/promoter replacement and various other applications. Here we combine the power of selectable markers followed by CRISPR/CAS9 genome editing for common genetic manipulations in yeast in a marker-free manner. We demonstrate our approach for whole gene and promoter replacements and for high-efficiency operator array integration. Our approach allows the utilization of many thousands of existing strains including library strains for the generation of significant genetic changes in yeast in a marker-free and cloning-free fashion.

引用

页码：1129 / 1139

页数：11

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