Gene activation in human cells using CRISPR/Cpf1-p300 and CRISPR/Cpf1-SunTag systems

Zhang, Xin; Wang, Wei; Shan, Lin; Han, Le; Ma, Shufeng; Zhang, Yan; Hao, Bingtao; Lin, Ying; Rong, Zhili

doi:10.1007/s13238-017-0491-6

Cited by 49 publications

(38 citation statements)

References 15 publications

(19 reference statements)

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“…By targeting the promoter region or the enhancer, gene expression could be enhanced. However, significant effects could only be obtained for dLbCas12a-p300, whereas dAsCas12a-p300 induced merely marginal activation [87]. Transcriptional repression mediated by dCas12a was demonstrated in plants with the use of dAsCas12a and dLbCas12a [74].…”

Section: Using Cas12a As a Dna Binding Proteinmentioning

confidence: 95%

“…An interesting aspect of target site cleavage by Cas12a is that apparently, cleavage of the nontarget strand is essential for cleavage of the target strand, a contrast to Cas9-mediated cleavage [84,85]. In human cells it was reported that dCas12abased transcription factors can be used for transcriptional activation, through either regulation on the transcriptional or epigenetic level [86,87]. Like dCas9, dCas12a can be used as a platform to recruit different enzymes to sites of interest (Fig.…”

Section: Using Cas12a As a Dna Binding Proteinmentioning

confidence: 99%

“…Like dCas9, dCas12a can be used as a platform to recruit different enzymes to sites of interest (Fig. However, significant effects could only be obtained for dLbCas12a-p300, whereas dAsCas12a-p300 induced merely marginal activation [87]. Currently, there are only few studies available showing the use of dCas12a as specific DNA binding protein.…”

Section: Using Cas12a As a Dna Binding Proteinmentioning

confidence: 99%

“…Currently, there are only few studies available showing the use of dCas12a as specific DNA binding protein. In human cells it was reported that dCas12abased transcription factors can be used for transcriptional activation, through either regulation on the transcriptional or epigenetic level [86,87]. By fusing a synthetic activator complex consisting of the VP16, p65, and Rta activator domains to dLbCas12a robust transcriptional activation could be achieved.…”

Section: Using Cas12a As a Dna Binding Proteinmentioning

confidence: 99%

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Transforming plant biology and breeding with CRISPR/Cas9, Cas12 and Cas13

2018

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Currently, biology is revolutionized by ever growing applications of the CRISPR/Cas system. As discussed in this Review, new avenues have opened up for plant research and breeding by the use of the sequence-specific DNases Cas9 and Cas12 (formerly named Cpf1) and, more recently, the RNase Cas13 (formerly named C2c2). Although double strand break-induced gene editing based on error-prone nonhomologous end joining has been applied to obtain new traits, such as powdery mildew resistance in wheat or improved pathogen resistance and increased yield in tomato, improved technologies based on CRISPR/Cas for programmed change in plant genomes via homologous recombination have recently been developed. Cas9- and Cas12- mediated DNA binding is used to develop tools for many useful applications, such as transcriptional regulation or fluorescence-based imaging of specific chromosomal loci in plant genomes. Cas13 has recently been applied to degrade mRNAs and combat viral RNA replication. By the possibility to address multiple sequences with different guide RNAs and by the simultaneous use of different Cas proteins in a single cell, we should soon be able to achieve complex changes of plant metabolism in a controlled way.

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Section: Using Cas12a As a Dna Binding Proteinmentioning

confidence: 95%

Section: Using Cas12a As a Dna Binding Proteinmentioning

confidence: 99%

Section: Using Cas12a As a Dna Binding Proteinmentioning

confidence: 99%

Section: Using Cas12a As a Dna Binding Proteinmentioning

confidence: 99%

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Transforming plant biology and breeding with CRISPR/Cas9, Cas12 and Cas13

2018

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“…BV3L6 Cas12a (dAsCas12a)-based VPR gene activator could be even more potent than dCas9-VPR (Liu et al 2017). It has been demonstrated in another study that the dLbCas12a-based SunTag system can also enable robust targeted gene activation (Zhang et al 2018b). Very recently, researchers have engineered an enhanced variant of AsCas12a termed enAsCas12a, which has a substantially expanded target range and improved editing efficiency (Kleinstiver et al 2019).…”

Section: Gene Gof By Dcas-tads In Animalsmentioning

confidence: 99%

The working dead: repurposing inactive CRISPR-associated nucleases as programmable transcriptional regulators in plants

Xiong

2019

aBIOTECH

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Targeted gene manipulation is highly desirable for fundamental plant research, plant synthetic biology, and molecular breeding. The clustered regularly interspaced short palindromic repeats-associated (Cas) nuclease is a revolutionary tool for genome editing, and has received snowballing popularity for gene knockout applications in diverse organisms including plants. Recently, the nuclease-dead Cas (dCas) proteins have been repurposed as programmable transcriptional regulators through translational fusion with portable transcriptional repression or activation domains, which has paved new ways for flexible and multiplex control over the activities of target genes of interest without the need to generate DNA lesions. Here, we review the most important breakthroughs of dCas transcriptional regulators in non-plant organisms and recent accomplishments of this growing field in plants. We also provide perspectives on future development directions of dCas transcriptional regulators in plant research in hope to stimulate their quick evolution and broad applications.

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iKA‐CRISPR hESCs for inducible and multiplex orthogonal gene knockout and activation

Sun

et al. 2018

FEBS Letters

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Human embryonic stem cells (hESCs) have a wide range of applications in early human embryonic development mimics, disease modeling, and cell therapy. To fulfill these applications, we established hESCs for inducible and multiplex orthogonal gene knockout and activation, which we named iKA-CRISPR hESCs. In cells, when complexed with a short guide RNA containing a 14-bp target sequence (14-bp gRNA) or a long 20-bp gRNA, the doxycycline-induced Cas9-p300 protein could activate gene transcription or cleave genomic DNA, respectively. We also demonstrate using iKA-CRISPR hESCs that knockout of OCT4 promoted differentiation, and developmentally relevant microRNAs and transcription factors could be efficiently activated. Thus, iKA-CRISPR hESCs provide a convenient platform to control gene expression networks and, therefore, facilitate the applications of hESCs in basic and translational biomedical research.

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Gene activation in human cells using CRISPR/Cpf1-p300 and CRISPR/Cpf1-SunTag systems

Cited by 49 publications

References 15 publications

Transforming plant biology and breeding with CRISPR/Cas9, Cas12 and Cas13

Transforming plant biology and breeding with CRISPR/Cas9, Cas12 and Cas13

The working dead: repurposing inactive CRISPR-associated nucleases as programmable transcriptional regulators in plants

iKA‐CRISPR hESCs for inducible and multiplex orthogonal gene knockout and activation

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