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 hei-tag: a highly efficient tag to boost targeted genome editing

2021 ◽  
Author(s):  
Thomas Thumberger ◽  
Tinatini Tavhelidse ◽  
Jose Arturo Gutierrez-Triana ◽  
Rebekka Medert ◽  
Alex Cornean ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Genome Editing ◽  
Nuclear Localization ◽  
High Efficiency ◽  
Basic Research ◽  
Model Systems ◽  
Nuclear Localization Signals ◽  
Highly Efficient ◽  
Cas9 Protein ◽  
Steric Hinderance

Precise, targeted genome editing by CRISPR/Cas9 is key for basic research and translational approaches in model and non-model systems. While active in all species tested so far, editing efficiencies still leave room for improvement. To reach its target, the bacterial Cas9 needs to be efficiently shuttled into the nucleus as attempted by fusion of nuclear localization signals (NLSs) to the Cas9 protein. Additional domains such as FLAG- or myc-tags are added for immediate detection or straight-forward purification. To avoid steric hinderance impacting on activity, amino acid linkers are employed connecting Cas9 and additional domains. We present the 'hei-tag (high efficiency-tag)', boosting the activity of the wide variety of CRISPR/Cas genome editing tools. The addition of the hei-tag to Cas9 or a C-to-T base editor dramatically enhances the respective targeting efficiency in model systems ranging from fish to mammals, including tissue culture applications. This allows to instantly upgrade existing and potentially highly adapted systems as well as establish novel highly efficient tools.

Maize tissue culture, transformation, and genome editing

2021 ◽  
Author(s):  
Albert P. Kausch ◽  
Kimberly Nelson-Vasilchik ◽  
Michael Tilelli ◽  
Joel P. Hague
Keyword(s):  
Tissue Culture ◽  
Genome Editing ◽  
Culture Transformation ◽  
Maize Tissue Culture

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.

Spontaneous production of a C-type rna virus in rat tissue culture lines

1973 ◽  
Vol 12 (3) ◽  
pp. 654-666 ◽  
Author(s):  
V. Klement ◽  
M. O. Nicolson ◽  
W. Nelson-Rees ◽  
R. V. Gilden ◽  
S. Oroszlan ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Rna Virus ◽  
Spontaneous Production ◽  
Rat Tissue

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 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
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