Targeted Transcriptional Activation in Plants Using a Potent Dead Cas9–Derived Synthetic Gene Activator

Li, Zhenxiang; Wang, Fengzhu; Li, Jianfeng

doi:10.1002/cpmb.89

Cited by 11 publications

(9 citation statements)

References 52 publications

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“…Our work suggests that the most effective target regions are within the 350 bp upstream of the TSS. Traditionally, gRNAs for dCas9-TV-based transcriptional activation are first optimized using plant protoplast systems (Li et al 2019b). Our findings that regions 350 bp upstream of the TSS are most effective for dCas9-TV-based transcriptional activation indicate that it can be productive by focusing on the small region without using the protoplast systems.…”

mentioning

confidence: 85%

Positional effects on efficiency of CRISPR/Cas9-based transcriptional activation in rice plants

et al. 2019

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The nuclease-dead Cas9 (dCas9) has been reprogrammed for transcriptional activation by fusing dCas9 to a transcriptional activation domain. In the presence of a guide RNA (gRNA), the dCas9 fusions specifically bind to regions of a promoter to activate transcription. Significant amount of effort has been directed toward the identification and optimization of the fusions of dCas9-activation domain, but very little is known about the impact of gRNA target positions within a promoter in plants on transcriptional activation efficiency. The dCas9-6TAL-VP128 system (dCas9-TV) has been optimized to activate transcription in plants. Here we use the dCas9-TV to activate transcription of OsWOX11 and OsYUC1, two genes that cause dramatic developmental phenotypes when overexpressed. We designed a series of gRNAs targeting the promoters of the two genes. We show that gRNAs that target regions within 350 bp upstream of the transcription start site were most effective in transcriptional activation. Moreover, we show that using two gRNAs that simultaneously target two discrete sites in a promoter can further enhance transcription. This work provides guidelines for designed transcriptional activation through CRISPR/dCas9 systems.

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confidence: 85%

Positional effects on efficiency of CRISPR/Cas9-based transcriptional activation in rice plants

et al. 2019

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“…This synthetic construct that is generated by tethering transcriptional activation domains to a dCas9 can be directed to the promoters of endogenous target genes by single guide RNAs (sgRNAs) to activate transcription. It can also be used to target various effector domains can be fused to dCas9 to enable programmable transcriptional and epigenetic control (Li et al, 2019).…”

Section: Cas 9 Systemmentioning

confidence: 99%

“…A number of researchers have utilized CRISPR systems to investigate the genetic targets behind differentiation blockages commonly seen in causing the development of AML. For instance, Wang et al, identified RNA-binding protein ZFP36L2 as a crucial mechanistic regulator of cell differentiation in AML using cell surface antigen-guided-CRISPR systems (Li et al, 2019;Wang et al, 2021a). Genetic suppression of ZFP36L2 enables myeloid leukemic cells to transit out from an undifferentiated state and resume cellular differentiations.…”

Section: Application Of Various Crispr System In Studying the Functional Genomics Of Amlmentioning

confidence: 99%

Utilization of CRISPR-Mediated Tools for Studying Functional Genomics in Hematological Malignancies: An Overview on the Current Perspectives, Challenges, and Clinical Implications

et al. 2022

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Hematological malignancies (HM) are a group of neoplastic diseases that are usually heterogenous in nature due to the complex underlying genetic aberrations in which collaborating mutations enable cells to evade checkpoints that normally safeguard it against DNA damage and other disruptions of healthy cell growth. Research regarding chromosomal structural rearrangements and alterations, gene mutations, and functionality are currently being carried out to understand the genomics of these abnormalities. It is also becoming more evident that cross talk between the functional changes in transcription and proteins gives the characteristics of the disease although specific mutations may induce unique phenotypes. Functional genomics is vital in this aspect as it measures the complete genetic change in cancerous cells and seeks to integrate the dynamic changes in these networks to elucidate various cancer phenotypes. The advent of CRISPR technology has indeed provided a superfluity of benefits to mankind, as this versatile technology enables DNA editing in the genome. The CRISPR-Cas9 system is a precise genome editing tool, and it has revolutionized methodologies in the field of hematology. Currently, there are various CRISPR systems that are used to perform robust site-specific gene editing to study HM. Furthermore, experimental approaches that are based on CRISPR technology have created promising tools for developing effective hematological therapeutics. Therefore, this review will focus on diverse applications of CRISPR-based gene-editing tools in HM and its potential future trajectory. Collectively, this review will demonstrate the key roles of different CRISPR systems that are being used in HM, and the literature will be a representation of a critical step toward further understanding the biology of HM and the development of potential therapeutic approaches.

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“…Controlling gene expression has long been a goal of plant scientists for deciphering plant gene function and improving crop traits. CRISPR-mediated gene regulation, designed by engineering a nuclease-inactivated Cas9 (dCas9) protein with a transcriptional activation domain (VP64) or repressor domains (SRDX), has been very powerful in activating or repressing the endogenous expression of plant genes of interest (Lowder et al, 2017;Li Z. et al, 2019;Malzahn et al, 2019). With the help of gRNA, the dCas9 fusion protein can specifically bind to the promoter region and manipulate the expression of downstream genes.…”

Section: Transcriptional Regulationmentioning

confidence: 99%

Multiplex Genome-Editing Technologies for Revolutionizing Plant Biology and Crop Improvement

et al. 2021

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Multiplex genome-editing (MGE) technologies are recently developed versatile bioengineering tools for modifying two or more specific DNA loci in a genome with high precision. These genome-editing tools have greatly increased the feasibility of introducing desired changes at multiple nucleotide levels into a target genome. In particular, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) [CRISPR/Cas] system-based MGE tools allow the simultaneous generation of direct mutations precisely at multiple loci in a gene or multiple genes. MGE is enhancing the field of plant molecular biology and providing capabilities for revolutionizing modern crop-breeding methods as it was virtually impossible to edit genomes so precisely at the single base-pair level with prior genome-editing tools, such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). Recently, researchers have not only started using MGE tools to advance genome-editing applications in certain plant science fields but also have attempted to decipher and answer basic questions related to plant biology. In this review, we discuss the current progress that has been made toward the development and utilization of MGE tools with an emphasis on the improvements in plant biology after the discovery of CRISPR/Cas9. Furthermore, the most recent advancements involving CRISPR/Cas applications for editing multiple loci or genes are described. Finally, insights into the strengths and importance of MGE technology in advancing crop-improvement programs are presented.

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Targeted Transcriptional Activation in Plants Using a Potent Dead Cas9–Derived Synthetic Gene Activator

Cited by 11 publications

References 52 publications

Positional effects on efficiency of CRISPR/Cas9-based transcriptional activation in rice plants

Positional effects on efficiency of CRISPR/Cas9-based transcriptional activation in rice plants

Utilization of CRISPR-Mediated Tools for Studying Functional Genomics in Hematological Malignancies: An Overview on the Current Perspectives, Challenges, and Clinical Implications

Multiplex Genome-Editing Technologies for Revolutionizing Plant Biology and Crop Improvement

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