scholarly journals Development of a pyrF-based counterselectable system for targeted gene deletion in Streptomyces rimosus

2021 ◽  
Vol 22 (5) ◽  
pp. 383-396
Author(s):  
Yiying Yang ◽  
Qingqing Sun ◽  
Yang Liu ◽  
Hanzhi Yin ◽  
Wenping Yang ◽  
...  
Keyword(s):  
Gene Deletion ◽  
Streptomyces Rimosus ◽  
Targeted Gene Deletion

scholarly journals Versatile CRISPR/Cas9 Systems for Genome Editing in Ustilago maydis

2021 ◽  
Vol 7 (2) ◽  
pp. 149
Author(s):  
Sarah-Maria Wege ◽  
Katharina Gejer ◽  
Fabienne Becker ◽  
Michael Bölker ◽  
Johannes Freitag ◽  
...  
Keyword(s):  
Genome Editing ◽  
Cell Biology ◽  
Gene Deletion ◽  
Fluorescent Protein ◽  
Point Mutations ◽  
Model Organism ◽  
Ustilago Maydis ◽  
Genomic Locus ◽  
Targeted Gene Deletion ◽  
Molecular Cell Biology

The phytopathogenic smut fungus Ustilago maydis is a versatile model organism to study plant pathology, fungal genetics, and molecular cell biology. Here, we report several strategies to manipulate the genome of U. maydis by the CRISPR/Cas9 technology. These include targeted gene deletion via homologous recombination of short double-stranded oligonucleotides, introduction of point mutations, heterologous complementation at the genomic locus, and endogenous N-terminal tagging with the fluorescent protein mCherry. All applications are independent of a permanent selectable marker and only require transient expression of the endonuclease Cas9hf and sgRNA. The techniques presented here are likely to accelerate research in the U. maydis community but can also act as a template for genome editing in other important fungi.

Targeted gene deletion in Brettanomyces bruxellensis with an expression-free CRISPR-Cas9 system

2020 ◽  
Vol 104 (16) ◽  
pp. 7105-7115
Author(s):  
Cristian Varela ◽  
Caroline Bartel ◽  
Cristobal Onetto ◽  
Anthony Borneman
Keyword(s):  
Gene Deletion ◽  
Brettanomyces Bruxellensis ◽  
Targeted Gene Deletion

Targeted gene deletion of Leishmania major genes encoding developmental stage-specific leishmanolysin (GP63)

1998 ◽  
Vol 27 (3) ◽  
pp. 519-530 ◽  
Author(s):  
Phalgun B. Joshi ◽  
David L. Sacks ◽  
Govind Modi ◽  
W. Robert McMaster
Keyword(s):  
Developmental Stage ◽  
Gene Deletion ◽  
Leishmania Major ◽  
Major Genes ◽  
Targeted Gene Deletion ◽  
Genes Encoding

Gene deletion suggests a role for Trypanosoma cruzi surface glycoprotein GP72 in the insect and mammalian stages of the life cycle

1993 ◽  
Vol 106 (4) ◽  
pp. 1023-1033 ◽  
Author(s):  
A.R. de Jesus ◽  
R. Cooper ◽  
M. Espinosa ◽  
J.E. Gomes ◽  
E.S. Garcia ◽  
...  
Keyword(s):  
Life Cycle ◽  
Trypanosoma Cruzi ◽  
Gene Deletion ◽  
Surface Glycoprotein ◽  
Triatoma Infestans ◽  
Insect Vector ◽  
Null Mutant ◽  
Apparent Lack ◽  
Targeted Gene Deletion ◽  
Mouse Macrophages

We have explored the biological function of a surface glycoprotein (GP72) of Trypanosoma cruzi by studying a null mutant parasite, generated by targeted gene deletion. GP72 deletion affected parasite morphology in several stages of the life cycle. Insect midgut (epimastigote) forms had a detached flagellum (apomastigote) in the null mutant. The abnormal flagellar phenotype persisted during development of the infective (metacyclic) forms but there was no impairment in the acquisition of complement resistance, sialidase expression or cell infectivity. The GP72 null mutant could efficiently infect and proliferate in mouse macrophages and non-phagocytic L6E9 cells. The mammalian stages of the life cycle also showed major morphological abnormalities. During early subcultures in L6E9 cells, few extracellular fully flagellated forms, expressing markers characteristic of trypomastigotes, were seen. The extracellular population consisted almost exclusively of rounded forms with short flagella (micromastigote), which expressed an amastigote-specific surface marker and no sialidase. The propagation of the parasite was not affected, despite the apparent lack of the trypomastigote forms, which are thought to be primarily responsible for cell invasion. After some subcultures, the extracellular population changed to about equal numbers of micromastigotes and a range of flagellated forms that still did not include true trypomastigotes. Instead, the kinetoplast remained close to the nucleus and the flagellum emerged from the middle of the cell (mesomastigote). Half of the flagellum adhered to the cell body and the remainder was free at the anterior end. In Triatoma infestans, the survival of the mutant was dramatically reduced, suggesting that either GP72 itself, or the altered properties of the flagellum, were critical for establishment in the insect vector.

scholarly journals Targeted gene deletion of heme oxygenase 2 reveals neural role for carbon monoxide

1997 ◽  
Vol 94 (26) ◽  
pp. 14848-14853 ◽  
Author(s):  
R. Zakhary ◽  
K. D. Poss ◽  
S. R. Jaffrey ◽  
C. D. Ferris ◽  
S. Tonegawa ◽  
...  
Keyword(s):  
Carbon Monoxide ◽  
Heme Oxygenase ◽  
Gene Deletion ◽  
Targeted Gene Deletion

Targeted Gene Deletion Using DNA-Free RNA-Guided Cas9 Nuclease Accelerates Adaptation of CHO Cells to Suspension Culture

2016 ◽  
Vol 5 (11) ◽  
pp. 1211-1219 ◽  
Author(s):  
Namil Lee ◽  
JongOh Shin ◽  
Jin Hyoung Park ◽  
Gyun Min Lee ◽  
Suhyung Cho ◽  
...  
Keyword(s):  
Suspension Culture ◽  
Cho Cells ◽  
Gene Deletion ◽  
Targeted Gene Deletion ◽  
Cas9 Nuclease

Targeted gene deletion inZygosaccharomyces bailii

Yeast ◽  
2001 ◽  
Vol 18 (2) ◽  
pp. 173-186 ◽  
Author(s):  
Mehdi Mollapour ◽  
Peter W. Piper
Keyword(s):  
Gene Deletion ◽  
Targeted Gene Deletion

scholarly journals Analysis of Noncanonical Calcium-Dependent Protein Kinases in Toxoplasma gondii by Targeted Gene Deletion Using CRISPR/Cas9

2016 ◽  
Vol 84 (5) ◽  
pp. 1262-1273 ◽  
Author(s):  
Shaojun Long ◽  
Qiuling Wang ◽  
L. David Sibley
Keyword(s):  
Toxoplasma Gondii ◽  
Protein Kinases ◽  
Gene Deletion ◽  
Cyst Formation ◽  
Content Type ◽  
Calcium Dependent ◽  
Targeted Gene Deletion ◽  
Dependent Protein ◽  
Calcium Dependent Protein Kinases

Calcium-dependent protein kinases (CDPKs) are expanded in apicomplexan parasites, especially inToxoplasma gondiiwhere 14 separate genes encoding these enzymes are found. Although previous studies have shown that several CDPKs play a role in controlling invasion, egress, and cell division inT. gondii, the roles of most of these genes are unexplored. Here we developed a more efficient method for gene disruption using CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) that was modified to completely delete large, multiexonic genes from the genome and to allow serial replacement by recycling of the selectable marker using Cre-loxP. Using this system, we generated a total of 24 mutants in type 1 and 2 genetic backgrounds to ascertain the functions of noncanonical CDPKs. Remarkably, although we were able to confirm the essentiality of CDPK1 and CDPK7, the majority of CDPKs had no discernible phenotype for growthin vitroor infection in the mouse model. The exception to this was CDPK6, loss of which leads to reduced plaquing, fitness defect in a competition assay, and reduced tissue cyst formation in chronically infected mice. Our findings highlight the utility of CRISPR/Cas9 for rapid serial gene deletion and also suggest that additional models are needed to reveal the functions of many genes inT. gondii.

scholarly journals Targeted Gene Deletion ofLeishmania majorUDP-galactopyranose Mutase Leads to Attenuated Virulence

2007 ◽  
Vol 282 (14) ◽  
pp. 10498-10505 ◽  
Author(s):  
Barbara Kleczka ◽  
Anne-Christin Lamerz ◽  
Ger van Zandbergen ◽  
Alexander Wenzel ◽  
Rita Gerardy-Schahn ◽  
...  
Keyword(s):  
Gene Deletion ◽  
Targeted Gene Deletion
Sign in / Sign up

Export Citation Format

Share Document