scholarly journals Cas9 cleavage assay for pre-screening of sgRNAs using nicking triggered isothermal amplification

2016 ◽  
Vol 7 (8) ◽  
pp. 4951-4957 ◽  
Author(s):  
Kaixiang Zhang ◽  
Ruijie Deng ◽  
Yue Li ◽  
Ling Zhang ◽  
Jinghong Li
Keyword(s):  
Genome Editing ◽  
Quantitative Evaluation ◽  
Isothermal Amplification ◽  
Highly Efficient ◽  
Cleavage Efficiency ◽  
Cleavage Assay

A novel Cas9 cleavage assay was developed for quantitative evaluation of Cas9 cleavage efficiency and pre-screening of sgRNA to achieve highly specific and highly efficient CRISPR/Cas9 genome editing.

scholarly journals An Efficient, Rapid, and Recyclable System for CRISPR-Mediated Genome Editing in Candida albicans

mSphere ◽  
2017 ◽  
Vol 2 (2) ◽  
Author(s):  
Namkha Nguyen ◽  
Morgan M. F. Quail ◽  
Aaron D. Hernday
Keyword(s):  
Candida Albicans ◽  
Genome Editing ◽  
Fungal Pathogen ◽  
Gene Knockout ◽  
Strain Engineering ◽  
Molecular Genetic Analysis ◽  
Content Type ◽  
Highly Efficient ◽  
Discovery Research ◽  
Gene Knockouts

ABSTRACT Candida albicans is the most common fungal pathogen of humans. Historically, molecular genetic analysis of this important pathogen has been hampered by the lack of stable plasmids or meiotic cell division, limited selectable markers, and inefficient methods for generating gene knockouts. The recent development of clustered regularly interspaced short palindromic repeat(s) (CRISPR)-based tools for use with C. albicans has opened the door to more efficient genome editing; however, previously reported systems have specific limitations. We report the development of an optimized CRISPR-based genome editing system for use with C. albicans. Our system is highly efficient, does not require molecular cloning, does not leave permanent markers in the genome, and supports rapid, precise genome editing in C. albicans. We also demonstrate the utility of our system for generating two independent homozygous gene knockouts in a single transformation and present a method for generating homozygous wild-type gene addbacks at the native locus. Furthermore, each step of our protocol is compatible with high-throughput strain engineering approaches, thus opening the door to the generation of a complete C. albicans gene knockout library. IMPORTANCE Candida albicans is the major fungal pathogen of humans and is the subject of intense biomedical and discovery research. Until recently, the pace of research in this field has been hampered by the lack of efficient methods for genome editing. We report the development of a highly efficient and flexible genome editing system for use with C. albicans. This system improves upon previously published C. albicans CRISPR systems and enables rapid, precise genome editing without the use of permanent markers. This new tool kit promises to expedite the pace of research on this important fungal pathogen.

Highly efficient heritable genome editing in wheat using an RNA virus and bypassing tissue culture

2021 ◽  
Author(s):  
Tingdong Li ◽  
Jiacheng Hu ◽  
Yu Sun ◽  
Boshu Li ◽  
Dingliang Zhang ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Genome Editing ◽  
Rna Virus ◽  
Highly Efficient

scholarly journals Highly Efficient Mouse Genome Editing by CRISPR Ribonucleoprotein Electroporation of Zygotes

2016 ◽  
Vol 291 (28) ◽  
pp. 14457-14467 ◽  
Author(s):  
Sean Chen ◽  
Benjamin Lee ◽  
Angus Yiu-Fai Lee ◽  
Andrew J. Modzelewski ◽  
Lin He
Keyword(s):  
Genome Editing ◽  
Mouse Genome ◽  
Highly Efficient

scholarly journals Optimized Cas9 expression systems for highly efficient Arabidopsis genome editing facilitate isolation of complex alleles in a single generation

2019 ◽  
Vol 20 (1) ◽  
pp. 151-162 ◽  
Author(s):  
Jana Ordon ◽  
Mauro Bressan ◽  
Carola Kretschmer ◽  
Luca Dall’Osto ◽  
Sylvestre Marillonnet ◽  
...  
Keyword(s):  
Genome Editing ◽  
Arabidopsis Genome ◽  
Expression Systems ◽  
Single Generation ◽  
Highly Efficient

Highly efficient genome editing in N. gerenzanensis using an inducible CRISPR/Cas9–RecA system

2020 ◽  
Vol 42 (9) ◽  
pp. 1699-1706
Author(s):  
Xue Yue ◽  
Tianyu Xia ◽  
Shuai Wang ◽  
Huijun Dong ◽  
Yongquan Li
Keyword(s):  
Genome Editing ◽  
Highly Efficient

scholarly journals Development of a Highly Efficient Multiplex Genome Editing System in Outcrossing Tetraploid Alfalfa (Medicago sativa)

2020 ◽  
Vol 11 ◽  
Author(s):  
Tezera W. Wolabu ◽  
Lili Cong ◽  
Jong-Jin Park ◽  
Qinyan Bao ◽  
Miao Chen ◽  
...  
Keyword(s):  
Medicago Sativa ◽  
Genome Editing ◽  
Highly Efficient
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