Faculty Opinions recommendation of Optimal design and analysis of genetic studies on gene expression.

ABSTRACT We developed a series of ligand-inducible riboswitches that control gene expression in diverse species of Gram-negative and Gram-positive bacteria, including human pathogens that have few or no previously reported inducible expression systems. We anticipate that these riboswitches will be useful tools for genetic studies in a wide range of bacteria.

Download Full-text

Two related families of retrotransposons from Schizosaccharomyces pombe.

Molecular and Cellular Biology ◽

10.1128/mcb.10.12.6791 ◽

1990 ◽

Vol 10 (12) ◽

pp. 6791-6798 ◽

Cited By ~ 68

Author(s):

H L Levin ◽

D C Weaver ◽

J D Boeke

Keyword(s):

Gene Expression ◽

Reverse Transcriptase ◽

Schizosaccharomyces Pombe ◽

Fission Yeast ◽

Protein Sequences ◽

Complete Sequence ◽

Long Terminal Repeats ◽

Genetic Studies ◽

Its Gene ◽

Repeated Dnas

Two related families of transposons were isolated from schizosaccharomyces pombe, an organism which has been the object of extensive genetic studies which had previously produced no evidence for the existence of such elements. These two classes of repeated DNAs, dubbed Tf1 (transposon of fission yeast 1) and Tf2 have many properties of retrotransposons. Tf1 and Tf2 both possess long terminal repeats and predicted protein sequences that resemble the protease, reverse transcriptase, and integrase domains of retroviruses. The chromosomal locations and total numbers of Tf1 and Tf2 differ greatly in various isolates of S. pombe. The Tf elements are expressed in the form of 4.5-kb mRNAs. The complete sequence of Tf1 was determined and suggests that a novel mechanism for regulating its gene expression may be used.

Download Full-text

Investigating Post-translational Modifications in Neuropsychiatric Disease: The Next Frontier in Human Post-mortem Brain Research

Frontiers in Molecular Neuroscience ◽

10.3389/fnmol.2021.689495 ◽

2021 ◽

Vol 14 ◽

Author(s):

Melanie J. Grubisha ◽

Robert A. Sweet ◽

Matthew L. MacDonald

Keyword(s):

Gene Expression ◽

Brain Research ◽

Research Direction ◽

Psychiatric Disease ◽

Post Mortem ◽

Genetic Studies ◽

Post Translational Modifications ◽

Neuropsychiatric Disease ◽

Post Mortem Brain ◽

Circuit Function

Gene expression and translation have been extensively studied in human post-mortem brain tissue from subjects with psychiatric disease. Post-translational modifications (PTMs) have received less attention despite their implication by unbiased genetic studies and importance in regulating neuronal and circuit function. Here we review the rationale for studying PTMs in psychiatric disease, recent findings in human post-mortem tissue, the required controls for these types of studies, and highlight the emerging mass spectrometry approaches transforming this research direction.

Download Full-text

A CRISPR/dCas9 toolkit for functional analysis of maize genes

Plant Methods ◽

10.1186/s13007-020-00675-5 ◽

2020 ◽

Vol 16 (1) ◽

Cited By ~ 1

Author(s):

Irene N. Gentzel ◽

Chan Ho Park ◽

Maria Bellizzi ◽

Guiqing Xiao ◽

Kiran R. Gadhave ◽

...

Keyword(s):

Gene Expression ◽

Functional Analysis ◽

Functional Genomics ◽

Protoplast Isolation ◽

Genetic Studies ◽

Transfection Method ◽

Important Trait ◽

Maize Gene

Abstract Background The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system has become a powerful tool for functional genomics in plants. The RNA-guided nuclease can be used to not only generate precise genomic mutations, but also to manipulate gene expression when present as a deactivated protein (dCas9). Results In this study, we describe a vector toolkit for analyzing dCas9-mediated activation (CRISPRa) or inactivation (CRISPRi) of gene expression in maize protoplasts. An improved maize protoplast isolation and transfection method is presented, as well as a description of dCas9 vectors to enhance or repress maize gene expression. Conclusions We anticipate that this maize protoplast toolkit will streamline the analysis of gRNA candidates and facilitate genetic studies of important trait genes in this transformation-recalcitrant plant.

Download Full-text

CRISPR/Cas9 Mediated-Disruption of ZNF543 Gene: An Approach Towards Discovering Its Relation to TRIM28 Gene in Parkinson's Disease

10.21203/rs.3.rs-797710/v1 ◽

2021 ◽

Author(s):

Mohammad Hashemabadi ◽

Hosseinali Sasan ◽

Mojdeh Amandadi ◽

Keyvan Esmaeilzadeh-Salestani ◽

Saeed Esmaeili-Mahani ◽

...

Keyword(s):

Gene Expression ◽

Parkinson’S Disease ◽

Parkinson's Disease ◽

Neuroblastoma Cell ◽

Regulatory Region ◽

Gene Mutations ◽

Neuroblastoma Cell Line ◽

Human Neuroblastoma Cell ◽

Human Neuroblastoma ◽

Genetic Studies

Abstract Genetic studies of familial forms of Parkinson’s disease (PD) have shown that the ZNF543 gene is a candidate gene that operates relevant to this disease. However, until now there is no evidence for ZNF543 gene function in PD, and mechanisms resulting from its mutation had not been elucidated. Given the same genetic location of the ZNF543 gene with TRIM28 and effects of both of them on PD pathogenesis, we surmised that ZNF543 may act as a transcription factor for TRIM28 gene expression. By knocking out the ZNF543 gene via the CRISPR/Cas9 editing platform, we assessed the functional effect of loss of expression of this gene on TRIM28 gene expression. Four sgRNAs with different PAM sequences were designed against two parts of the regulatory region of ZNF543 gene, and highly efficient disruption of ZNF543 expression in human neuroblastoma cell line was evaluated by polymerase chain reaction and T7 endonuclease assay. Moreover, evaluation of TRIM28 gene expression in ZNF543-knocked-out cells indicated a significant increase of TRIM28 gene expression, suggesting that ZNF543 probably regulates the expression of TRIM28. This approach offers a window into pinpointing the mechanism by which ZNF543 gene mutations mediate PD pathogenicity.

Download Full-text

Two related families of retrotransposons from Schizosaccharomyces pombe

Molecular and Cellular Biology ◽

10.1128/mcb.10.12.6791-6798.1990 ◽

1990 ◽

Vol 10 (12) ◽

pp. 6791-6798

Author(s):

H L Levin ◽

D C Weaver ◽

J D Boeke

Keyword(s):

Gene Expression ◽

Reverse Transcriptase ◽

Schizosaccharomyces Pombe ◽

Fission Yeast ◽

Protein Sequences ◽

Complete Sequence ◽

Long Terminal Repeats ◽

Genetic Studies ◽

Its Gene ◽

Repeated Dnas

Two related families of transposons were isolated from schizosaccharomyces pombe, an organism which has been the object of extensive genetic studies which had previously produced no evidence for the existence of such elements. These two classes of repeated DNAs, dubbed Tf1 (transposon of fission yeast 1) and Tf2 have many properties of retrotransposons. Tf1 and Tf2 both possess long terminal repeats and predicted protein sequences that resemble the protease, reverse transcriptase, and integrase domains of retroviruses. The chromosomal locations and total numbers of Tf1 and Tf2 differ greatly in various isolates of S. pombe. The Tf elements are expressed in the form of 4.5-kb mRNAs. The complete sequence of Tf1 was determined and suggests that a novel mechanism for regulating its gene expression may be used.

Download Full-text

Pollen Transcriptome and Proteome: Molecular and Functional Analysis

Advances in Cell Biology ◽

10.2478/v10052-010-0003-9 ◽

2010 ◽

Vol 2 (1) ◽

pp. 29-57 ◽

Cited By ~ 2

Author(s):

Katarzyna Rafinska ◽

Krzysztof Zienkiewicz ◽

Elzbieta Bednarska

Keyword(s):

Gene Expression ◽

Functional Analysis ◽

Structural Organization ◽

Current Knowledge ◽

Male Gametophyte ◽

Plant Cells ◽

Pollen Grain ◽

Rapid Progress ◽

Genetic Studies ◽

Simple Extraction

SummaryThe pollen grain, because of its unique structural organization, is an extremely useful experimental model in cytological, molecular as well as in genetic studies. Due to the ease of pollen grain isolation, their sorting as well as simple extraction of their DNA, RNA and proteins, male gametophyte cells of angiosperms are presently one of the most intensively studied plant cells. Important and rapid progress in the development of experimental tools for genome exploration caused a significant increase in the number of reports concerning different aspects of gene expression during microsporogenesis and microgametogenesis in angiosperm plants. In this review we present the current knowledge of the pollen transcriptome and proteome during different stages of male gametophyte development, especially in Arabidopsis thaliana. Most of the results presented here were obtained in experiments carried out using microarrays, which were designed on the basis of the known sequence of the Arabidopsis genome.

Download Full-text

Establishment of a Gene Expression System in Ochrobactrum anthropi

Applied and Environmental Microbiology ◽

10.1128/aem.01446-06 ◽

2006 ◽

Vol 72 (10) ◽

pp. 6833-6836 ◽

Cited By ~ 8

Author(s):

Mohamed N. Seleem ◽

Mohammed Ali ◽

Stephen M. Boyle ◽

Biswarup Mukhopadhyay ◽

Sharon G. Witonsky ◽

...

Keyword(s):

Gene Expression ◽

Genetic Manipulation ◽

Expression System ◽

Protein Detection ◽

Ochrobactrum Anthropi ◽

Genetic Studies ◽

Gene Expression System ◽

His Tag ◽

N Terminus

ABSTRACT Genetic studies of Ochrobactrum anthropi are hindered by the lack of a suitable gene expression system. We constructed a set of vectors containing several promoters and a His tag fusion in the N terminus to facilitate protein detection and purification. The new vectors should significantly enhance the genetic manipulation and characterization of O. anthropi.

Download Full-text