TALEN- and CRISPR-enhanced DNA homologous recombination for gene editing in zebrafish

2016 ◽  
pp. 107-120 ◽  
Author(s):  
Y. Zhang ◽  
H. Huang ◽  
B. Zhang ◽  
S. Lin
Keyword(s):  
Homologous Recombination ◽  
Gene Editing

scholarly journals Unsuccessful attempt at gene-editing by homologous recombination in the zebrafish germ line using the approach of “Rong and Golic”

2012 ◽  
Vol 21 (5) ◽  
pp. 1125-1136 ◽  
Author(s):  
Rosalind Brookfield ◽  
Felix Dafhnis-Calas ◽  
Zhengyao Xu ◽  
William Brown
Keyword(s):  
Homologous Recombination ◽  
Gene Editing ◽  
Germ Line ◽  
Unsuccessful Attempt

scholarly journals Evaluation of site-specific homologous recombination activity of BRCA1 by direct quantitation of gene editing efficiency

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Yuki Yoshino ◽  
Shino Endo ◽  
Zhenghao Chen ◽  
Huicheng Qi ◽  
Gou Watanabe ◽  
...  
Keyword(s):  
Homologous Recombination ◽  
Gene Editing ◽  
Recombination Activity ◽  
Site Specific ◽  
Editing Efficiency

PML isoform expression and DNA break location relative to PML nuclear bodies impacts the efficiency of homologous recombination

2020 ◽  
Vol 98 (3) ◽  
pp. 314-326 ◽  
Author(s):  
Kathleen M. Attwood ◽  
Jayme Salsman ◽  
Dudley Chung ◽  
Sabateeshan Mathavarajah ◽  
Carter Van Iderstine ◽  
...  
Keyword(s):  
Homologous Recombination ◽  
Gene Editing ◽  
Genotoxic Stress ◽  
Nuclear Bodies ◽  
Homology Directed Repair ◽  
Pml Nuclear Bodies ◽  
Promyelocytic Leukemia Nuclear Bodies ◽  
Chromosomal Break ◽  
The Impact

Promyelocytic leukemia nuclear bodies (PML NBs) are nuclear subdomains that respond to genotoxic stress by increasing in number via changes in chromatin structure. However, the role of the PML protein and PML NBs in specific mechanisms of DNA repair has not been fully characterized. Here, we have directly examined the role of PML in homologous recombination (HR) using I-SceI extrachromosomal and chromosome-based homology-directed repair (HDR) assays, and in HDR by CRISPR/Cas9-mediated gene editing. We determined that PML loss can inhibit HR in an extrachromosomal HDR assay but had less of an effect on CRISPR/Cas9-mediated chromosomal HDR. Overexpression of PML also inhibited both CRISPR HDR and I-SceI-induced HDR using a chromosomal reporter, and in an isoform-specific manner. However, the impact of PML overexpression on the chromosomal HDR reporter was dependent on the intranuclear chromosomal positioning of the reporter. Specifically, HDR at the TAP1 gene locus, which is associated with PML NBs, was reduced compared with a locus not associated with a PML NB; yet, HDR could be reduced at the non-PML NB-associated locus by PML overexpression. Thus, both loss and overexpression of PML isoforms can inhibit HDR, and proximity of a chromosomal break to a PML NB can impact HDR efficiency.

Improvement of Chloroplast Transformation Using CRISPR/Cas9

2020 ◽  
Vol 14 (3) ◽  
pp. 401-407
Author(s):  
Ning Tang ◽  
Yumei Xia ◽  
Yijie Zhan ◽  
Junhao Dan ◽  
Mulan Yu ◽  
...  
Keyword(s):  
Dna Damage ◽  
Homologous Recombination ◽  
Gene Editing ◽  
Transformation Efficiency ◽  
Chloroplast Transformation ◽  
Nuclear Transformation ◽  
Important Research ◽  
Transformation System ◽  
Repair Mechanism ◽  
Damage Repair

Chloroplasts are organelles that contain genetic materials (DNA) in higher plant cells. The special genetic characteristics of chloroplasts mean that plasmid transformation has important research value, so it has become an important research direction second to nuclear transformation. Although the techniques of chloroplast genome modification have been successfully applied in tobacco and extended to other high plants, there are still many limitations. Exogenous genes are integrated into the chloroplast genome through homologous recombination. Therefore, the low efficiency of homologous recombination directly limits transformation efficiency. Gene editing with fixed-point cutting function and DNA damage repair mechanism may effectively improve the efficiency. In the present study, we aimed to use CRISPR/Cas9 to cut the site between two homologous recombinant fragments in chloroplast transformation to improve the efficiency by activating the DNA damage repair mechanism. The Cas9 gene and gRNA were added to the chloroplast transformation system of tobacco by co-transformation or integration into a transformation vector. The acquired resistant plants were screened by multiple selection of spectinomycin and chloroplast DNA was isolated for molecular detection by PCR. The results showed that the efficiency of chloroplast transformation increased by 6–10 times with the addition of gene editing technology. Although the transformation efficiency was still far below the level of nuclear transformation, this study may help to increase the efficiency of the plant chloroplast transformation system, and expand the types of plant receptors.

scholarly journals Gene Editing a Constitutively Active OsRac1 by Homologous Recombination-Based Gene Targeting Induces Immune Responses in Rice

2013 ◽  
Vol 54 (12) ◽  
pp. 2058-2070 ◽  
Author(s):  
Thu Thi Dang ◽  
Zenpei Shimatani ◽  
Yoji Kawano ◽  
Rie Terada ◽  
Ko Shimamoto
Keyword(s):  
Homologous Recombination ◽  
Immune Responses ◽  
Gene Targeting ◽  
Gene Editing ◽  
Constitutively Active

scholarly journals Germline Gene Editing in Chickens by Efficient CRISPR-Mediated Homologous Recombination in Primordial Germ Cells

PLoS ONE ◽  
2016 ◽  
Vol 11 (4) ◽  
pp. e0154303 ◽  
Author(s):  
Lazar Dimitrov ◽  
Darlene Pedersen ◽  
Kathryn H. Ching ◽  
Henry Yi ◽  
Ellen J. Collarini ◽  
...  
Keyword(s):  
Homologous Recombination ◽  
Germ Cells ◽  
Gene Editing ◽  
Primordial Germ Cells ◽  
Germline Gene

scholarly journals Corrigendum: Enhancement of Precise Gene Editing by the Association of Cas9 With Homologous Recombination Factors

2020 ◽  
Vol 11 ◽  
Author(s):  
Ngoc-Tung Tran ◽  
Sanum Bashir ◽  
Xun Li ◽  
Jana Rossius ◽  
Van Trung Chu ◽  
...  
Keyword(s):  
Homologous Recombination ◽  
Gene Editing

scholarly journals Gene editing in bacteria via SpCas9/sgRNA ribonucleoprotein complexes

2021 ◽  
Author(s):  
Ruben Dario Arroyo-Olarte ◽  
Ricardo Neftali Bravo-Rodriguez ◽  
Edgar Morales-Rios
Keyword(s):  
Green Fluorescent Protein ◽  
Homologous Recombination ◽  
Gene Editing ◽  
Fluorescent Protein ◽  
High Toxicity ◽  
Guide Rna ◽  
Ribonucleoprotein Complexes ◽  
Gfp Reporter ◽  
Blue Fluorescent Protein ◽  
Green Fluorescent

Gene editing has been revolutionized by CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas technology in a variety of organisms. In bacteria, however, CRISPR-Cas still holds many caveats such as high toxicity of Cas9 and off-target editing effects. In this work we develop a system for the incorporation of Cas9/single guide RNA ribonucleoprotein complexes in bacteria and their successful application for gene editing via homologous recombination. Targeting of a green fluorescent protein (GFP) reporter allows for easy verification of gene-edition via conversion to blue-fluorescent protein (BFP), mediated by the well characterized 196T > C (Tyr66His) mutation.

scholarly journals Development of an Efficient Gene Editing Tool in Schizochytrium sp. and Improving Its Lipid and Terpenoid Biosynthesis

2021 ◽  
Vol 8 ◽  
Author(s):  
Peng-Wei Huang ◽  
Ying-Shuang Xu ◽  
Xiao-Man Sun ◽  
Tian-Qiong Shi ◽  
Yang Gu ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Homologous Recombination ◽  
Gene Editing ◽  
Saturated Fatty Acids ◽  
Cell Factory ◽  
Terpenoid Biosynthesis ◽  
Acetyl Coa ◽  
Knock Out ◽  
Cell Factories ◽  
Efficient Gene

Schizochytrium sp. HX-308 is a marine microalga with fast growth and high lipid content, which has potential as microbial cell factories for lipid compound biosynthesis. It is significant to develop efficient genetic editing tool and discover molecular target in Schizochytrium sp. HX-308 for lipid compound biosynthesis. In this study, we developed an efficient gene editing tool in HX-308 which was mediated by Agrobacterium tumefaciens AGL-1. Results showed that the random integration efficiency reached 100%, and the homologous recombination efficiency reached about 30%. Furthermore, the metabolic pathway of lipid and terpenoid biosynthesis were engineered. Firstly, the acetyl-CoA c-acetyltransferase was overexpressed in HX-308 with a strong constitutive promoter. With the overexpression of acetyl-CoA c-acetyltransferase, more acetyl-CoA was used to synthesize terpenoids, and the production of squalene, β-carotene and astaxanthin was increased 5.4, 1.8, and 2.4 times, respectively. Interestingly, the production of saturated fatty acids and polyunsaturated fatty acids also changed. Moreover, three Acyl-CoA oxidase genes which catalyze the first step of β-oxidation were knocked out using homologous recombination. Results showed that the production of lipids increased in the three knock-out strains. Our results demonstrated that the A. tumefaciens-mediated transformation method will be of great use for the study of function genes, as well as developing Schizochytrium sp. as a strong cell factory for producing high value products.

Sign in / Sign up

Export Citation Format

Share Document