CRISPR-dependent endogenous gene regulation is required for virulence in piscine Streptococcus agalactiae

ABSTRACTClustered regularly interspaced palindromic repeats (CRISPR) and their associatedcasgenes have been demonstrated to regulate self-genes and virulence in many pathogens. In this study, we found that inactivation ofcas9caused reduced adhesion and intracellular survival of the piscineStreptococcus agalactiaestrain GD201008-001 and significantly decreased the virulence of this strain in zebrafish and mice. Further investigation indicated that theregRtranscriptional regulator was upregulated in the Δcas9mutant. AsregRmediates the repression of hyaluronidase, a critical factor involved in opening the blood-brain barrier (BBB) in mice,cas9-mediated repression ofregRtranscription is important forS. agalactiaeto open the BBB and thereby cause meningitis in animals. This study expands our understanding of endogenous gene regulation mediated by CRISPR-Cas systems in bacteria.

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Heritable endogenous gene regulation in plants with designed polydactyl zinc finger transcription factors

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.192412899 ◽

2002 ◽

Vol 99 (20) ◽

pp. 13296-13301 ◽

Cited By ~ 72

Author(s):

X. Guan ◽

J. Stege ◽

M. Kim ◽

Z. Dahmani ◽

N. Fan ◽

...

Keyword(s):

Gene Regulation ◽

Transcription Factors ◽

Zinc Finger ◽

Endogenous Gene

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Gene Regulation Systems for Gene Therapy Applications in the Central Nervous System

Neurology Research International ◽

10.1155/2012/595410 ◽

2012 ◽

Vol 2012 ◽

pp. 1-10 ◽

Cited By ~ 16

Author(s):

Jerusha Naidoo ◽

Deborah Young

Keyword(s):

Gene Therapy ◽

Central Nervous System ◽

Nervous System ◽

Gene Regulation ◽

Transgene Expression ◽

Endogenous Gene ◽

Temporal Regulation ◽

Regulatory Systems ◽

Substantial Progress ◽

The Central Nervous System

Substantial progress has been made in the development of novel gene therapy strategies for central nervous system (CNS) disorders in recent years. However, unregulated transgene expression is a significant issue limiting human applications due to the potential side effects from excessive levels of transgenic protein that indiscriminately affect both diseased and nondiseased cells. Gene regulation systems are a tool by which tight tissue-specific and temporal regulation of transgene expression may be achieved. This review covers the features of ideal regulatory systems and summarises the mechanics of current exogenous and endogenous gene regulation systems and their utility in the CNS.

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TriTag: an integrative tool to correlate chromatin dynamics and gene expression in living cells

Nucleic Acids Research ◽

10.1093/nar/gkaa906 ◽

2020 ◽

Vol 48 (22) ◽

pp. e127-e127 ◽

Cited By ~ 1

Author(s):

Haiyue Xu ◽

Junyan Wang ◽

Ying Liang ◽

Yujuan Fu ◽

Sihui Li ◽

...

Keyword(s):

Gene Expression ◽

Gene Regulation ◽

Fluorescent Protein ◽

Chromatin Dynamics ◽

Endogenous Gene ◽

Dna Labeling ◽

Functional Capabilities ◽

Protein Tracking ◽

Response To Stress ◽

Transcriptional Bursting

Abstract A wealth of single-cell imaging studies have contributed novel insights into chromatin organization and gene regulation. However, a comprehensive understanding of spatiotemporal gene regulation requires developing tools to combine multiple monitoring systems in a single study. Here, we report a versatile tag, termed TriTag, which integrates the functional capabilities of CRISPR-Tag (DNA labeling), MS2 aptamer (RNA imaging) and fluorescent protein (protein tracking). Using this tag, we correlate changes in chromatin dynamics with the progression of endogenous gene expression, by recording both transcriptional bursting and protein production. This strategy allows precise measurements of gene expression at single-allele resolution across the cell cycle or in response to stress. TriTag enables capturing an integrated picture of gene expression, thus providing a powerful tool to study transcriptional heterogeneity and regulation.

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Multiplexed sgRNA Expression Allows Versatile Single Nonrepetitive DNA Labeling and Endogenous Gene Regulation

ACS Synthetic Biology ◽

10.1021/acssynbio.7b00268 ◽

2017 ◽

Vol 7 (1) ◽

pp. 176-186 ◽

Cited By ~ 13

Author(s):

Shipeng Shao ◽

Lei Chang ◽

Yuao Sun ◽

Yingping Hou ◽

Xiaoying Fan ◽

...

Keyword(s):

Gene Regulation ◽

Endogenous Gene ◽

Dna Labeling

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511. Endogenous Gene Regulation with Polydactyl Zinc Finger Transcription Factors

Molecular Therapy ◽

10.1016/s1525-0016(16)43341-1 ◽

2002 ◽

Vol 5 (5) ◽

pp. S168-S169

Keyword(s):

Gene Regulation ◽

Transcription Factors ◽

Zinc Finger ◽

Endogenous Gene

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Multiplexed sgRNA Expression Allows Versatile Single Non-repetitive DNA Labeling and Endogenous Gene Regulation

10.1101/121905 ◽

2017 ◽

Cited By ~ 2

Author(s):

Shipeng Shao ◽

Lei Chang ◽

Yuao Sun ◽

Yingping Hou ◽

Xiaoying Fan ◽

...

Keyword(s):

Gene Regulation ◽

Significant Contribution ◽

Genome Structure ◽

Live Cells ◽

Expression Systems ◽

Genomic Locus ◽

Endogenous Gene ◽

Dna Labeling ◽

Pathological Conditions ◽

In Series

ABSTRACTThe CRISPR/Cas9 system has made significant contribution to genome editing, gene regulation and chromatin studies in recent years. High-throughput and systematic investigations into the multiplexed biological systems and disease conditions require simultaneous expression and coordinated functioning of multiple sgRNAs. However, current co-transfection based sgRNA co-expression systems remain poorly efficient and virus-based transfection approaches are relatively costly and labor intensive. Here we established a vector-independent method allowing multiple sgRNA expression cassettes to be assembled in series into a single plasmid. This synthetic biology-based strategy excels in its efficiency, controllability and scalability. Taking the flexibility advantage of this all-in-one sgRNA expressing system, we further explored its applications in single non-repetitive genomic locus imaging as well as coordinated gene regulation in live cells. With its strong potency, our method will greatly facilitate the understandings in genome structure, function and dynamics, and will contribute to the systemic investigations into complex physiological and pathological conditions.

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Engineering Magnetic Nanomaterial Production in Magnetotactic Bacteria Through Gene Regulation

ASME 2012 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2012-80446 ◽

2012 ◽

Author(s):

Mary E. Wilson ◽

Lina M. González ◽

Warren C. Ruder ◽

Philip R. LeDuc

Keyword(s):

Gene Regulation ◽

Magnetic Stimulation ◽

Gene Network ◽

Regulatory Networks ◽

Magnetotactic Bacteria ◽

Genetic Regulatory Networks ◽

Endogenous Gene ◽

Network Modules ◽

Magnetic Nanomaterial ◽

Magnetospirillum Magneticum

Magnetotactic bacteria endogenously synthesize intracellular magnetic nanoparticles (magnetosomes); however, little is known regarding the genetic regulatory networks that control magnetosome production. In this paper, we explore the genetic response of Magnetospirillum magneticum strain AMB-1 to an applied electromagnetic field as a means to identify genes activated by magnetic stimulation. The expression of magnetosome island, flagellar and cytoskeletal genes was found to be differentially altered by magnetic stimulation at short and long times points. These results indicate previously uncharacterized endogenous gene network modules that could be exploited to engineer magnetic bacteria as magnetic nanomaterial producing-machines through gene regulation.

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