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Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex
Open accessAbstract
Systematic interrogation of gene function requires the ability to perturb gene expression in a robust and generalizable manner. Here we describe structure-guided engineering of a CRISPR-Cas9 complex to mediate efficient transcriptional activation at endogenous genomic loci. We used these engineered Cas9 activation complexes to investigate single-guide RNA (sgRNA) targeting rules for effective transcriptional activation, to demonstrate multiplexed activation of ten genes simultaneously, and to upregulate long intergenic non-coding RNA (lincRNA) transcripts. We also synthesized a library consisting of 70,290 guides targeting all human RefSeq coding isoforms to screen for genes that, upon activation, confer resistance to a BRAF inhibitor. The top hits included genes previously shown to be able to confer resistance, and novel candidates were validated using individual sgRNA and complementary DNA overexpression. A gene expression signature based on the top screening hits correlated with markers of BRAF inhibitor resistance in cell lines and patient-derived samples. These results collectively demonstrate the potential of Cas9-based activators as a powerful genetic perturbation technology.
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“Because of the relatively large size of 4277 bp of dCas9-VPR cDNA, the authors used a dual rAAV system to express the dCas9-VPR and gRNA expression cassettes [223,224].”
From Recombinant Adeno-Associated Viral Vectors (rAAV)-Vector Elements in Ocular Gene Therapy Clinical Trials and Transgene Expression and Bioactivity Assays (2020) · cites this paper as [224]
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“For example, CRISPRi (interference)23–25 and CRISPRa (activation)26–30 technologies utilize catalytically inactive Cas9 (dCas9) fused to either a transcriptional repressor (e.g., KRAB, Krüppel associated box) or a transcriptional activator (e.g., VPR, VP64-p65-Rta).”
From Large-scale CRISPR screening in primary human 3D gastric organoids enables comprehensive dissection of gene-drug interactions (2025) · cites this paper as [30]
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“Novel strategies of Cas9-based transcription modulators rapidly appeared prompted by the fusion of dCas9 or gRNA to different activator or repressor elements, with multiple degrees of modulation and specificity, and enabling single or multiplexed transcriptional control [74,75,76].”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [74]
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“This dCas9-effector strategy has already been applied in genome wide screening study in which gain-of-function mutations in melanoma cells that confer drug resistance were identified.46”
From CRISPR-Cas9: a new and promising player in gene therapy (2015) · cites this paper as [46]
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BackgroundMethodInfluentialNext-generation CRISPR/Cas9 transcriptional activation in Drosophila using flySAM
Jia, Y., Xu, R., Ren, X., Ewen‐Campen, B., Rajakumar, R., Zirin, J. D., Yang‐Zhou, D., Zhu, R., Wang, F., Mao, D., et al. (2018) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1800677115 · Cited by 68
“In contrast to VPR, SAM involves two separate protein components—dCas9-VP64 and MCP:p65-HSF1—as well as a modified sgRNA containing two MS2 hairpin structures that recruit MCP:p65-HSF1 (9).”
BackgroundInfluentialDeaner, M., Mejia, J., & Alper, H. S. (2017) · ACS Synthetic Biology · doi:10.1021/acssynbio.7b00163 · Cited by 67
Micro-RNAs in the regulation of immune response against SARS CoV-2 and other viral infections
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Genome Editing in iPSC-Based Neural Systems: From Disease Models to Future Therapeutic Strategies
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“States CRISPRi/a tools allow the modulation of gene expression at the endogenous transcription level, an advantage compared to RNA interference (RNAi) and complementary DNA (cDNA) library screening approaches (Larson et al., 2013; Konermann et al., 2015).”
BackgroundThe determinants of alternative RNA splicing in human cells
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CRISPR Gene Editing in the Kidney
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“Compared with the traditional expression techniques that are based on the modification of genes and promoters, the CRISPR/dCas-mediated transcription activation method (CRISPRa) is simple, target-specific, programmable and frequently applicable [4, 6, 7].”
MethodRNA-targeting CRISPR systems from metagenomic discovery to transcriptomic engineering
Smargon, A. A., Shi, Y. J., & Yeo, G. W.-M. (2020) · Nature Cell Biology · doi:10.1038/s41556-019-0454-7 · Cited by 64
Engineering Metabolism in Nicotiana Species: A Promising Future
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“However, this comparison used initial and less effective CRISPR activators and not the most enhanced genetically engineered improvements [84, 87] that are described below.”
BackgroundPathophysiological Functions of Rnd 3/ RhoE
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Applying CRISPR–Cas9 tools to identify and characterize transcriptional enhancers
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“Inactivated Cas proteins can be used to deliver transcriptional regulator domains to the promoter of the gene of interest by direct fusion of regulatory domains, or repetitive peptide epitopes that recruit multiple copies of antibody-fused regulators (SunTag), or by using MS2 RNA stem-loops in the sequence of the trac…”
BackgroundDesigned nucleases for targeted genome editing
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Advances in detecting and reducing off-target effects generated by CRISPR-mediated genome editing
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“Up to ten targets including MYOD1, ASCL1, and KLF4 and lincRNA transcripts (Konermann et al., 2015)”
BackgroundFrequent sgRNA-barcode recombination in single-cell perturbation assays
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“Similarly, the synergistic activation mediator (SAM) system uses two MS2 hairpins in the sgRNA and fuses MCP to the activators p65 and HSF-1 (Heat Shock Factor 1, responsible for transcribing genes in response to temperature) [56].”
BackgroundCRISPR/Cas9 Epigenome Editing Potential for Rare Imprinting Diseases: A Review
Syding, L. A., Nickl, P., Kašpárek, P., & Sedláček, R. (2020) · Cells · doi:10.3390/cells9040993 · Cited by 65
“This scaffold can associate with dCas-V64 fusion protein and give rise to more efficient transcription activator [68,77].”
BackgroundCRISPR–Cas9-mediated genome editing and guide RNA design
Wiles, M. V., Qin, W., Cheng, A. W., & Wang, H. (2015) · Mammalian Genome · doi:10.1007/s00335-015-9565-z · Cited by 67
“Moreover, different stem loop structures recognized by RNA binding proteins have been engineered into the sgRNA backbone, which serve as a bait to recruit different effectors, for the purpose of gene activation and repression (Konermann et al. 2014; Zalatan et al. 2015).”
BackgroundInfluentialReversible Disruption of Specific Transcription Factor-DNA Interactions Using CRISPR/Cas9
Shariati, S. A., Dominguez, A. A., Xie, S., Wernig, M., Qi, L. S., & Skotheim, J. M. (2019) · Molecular Cell · doi:10.1016/j.molcel.2019.04.011 · Cited by 69
“Similarly, CRISPRa approaches to activate expression of specific genes by using dCas9 to recruit transcriptional activators does not determine the function of specific TF-DNA binding sites (Konermann et al., 2015; Tanenbaum etal., 2014).”
RNA-guided retargeting of Sleeping Beauty transposition in human cells
Kovač, A., Miskey, C., Menzel, M., Grueso, E., Gogol‐Döring, A., & Ivics, Z. (2020) · eLife · doi:10.7554/elife.53868 · Cited by 61
Hepatic stellate cell reprogramming via exosome-mediated CRISPR/dCas9-VP64 delivery
Luo, N., Li, J., Chen, Y., Xu, Y., Wei, Y., Lu, J., & Dong, R. (2020) · Drug Delivery · doi:10.1080/10717544.2020.1850917 · Cited by 64
“ogy can knock out, edit, inhibit and activate genes, and CRISPR/dCas9-VP64 system has an efficient gene transcriptional activation function in this study (Konermann et al., 2015).”
BackgroundMethodInfluentialThe Role of the Microbiota–Gut–Brain Axis and Antibiotics in ALS and Neurodegenerative Diseases
Obrenovich, M., Jaworski, H., Tadimalla, T., Mistry, A., Sykes, L., Perry, G., & Bonomo, R. (2020) · Microorganisms · doi:10.3390/microorganisms8050784 · Cited by 62
“Here, we review past and current methods used for the identification of FALS and SALS associated genes and propose a risk-based classification for these [41].”
BackgroundMethodLncRNAs in Cancer: From garbage to Junk
Aprile, M., Katopodi, V., Leucci, E., & Costa, V. (2020) · Cancers · doi:10.3390/cancers12113220 · Cited by 63
Murakami, K., Terakado, Y., Saito, K., Jomen, Y., Takeda, H., Oshima, M., & Barker, N. (2021) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2016806118 · Cited by 62
“4) to ensure that most cells receive only one viral construct with high probability (40).”
BackgroundDanziger, O., Patel, R. S., DeGrace, E. J., Rosen, M. R., & Rosenberg, B. R. (2022) · PLoS Pathogens · doi:10.1371/journal.ppat.1010464 · Cited by 62
“Advantages include easy to produce gRNA libraries, physiologically relevant expression levels, and multiplexing capabilities [42].”
BackgroundSynthetic biology and opportunities within agricultural crops
Sargent, D., Conaty, W. C., Tissue, D. T., & Sharwood, R. E. (2022) · Journal of Sustainable Agriculture and Environment · doi:10.1002/sae2.12014 · Cited by 63
KCNQ1OT1 promotes genome-wide transposon repression by guiding RNA–DNA triplexes and HP1 binding
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Application of CRISPR/Cas9 for biomedical discoveries
Riordan, S. M., Heruth, D. P., Zhang, L. Q., & Ye, S. Q. (2015) · Cell & Bioscience · doi:10.1186/s13578-015-0027-9 · Cited by 66
“These techniques either induce random indels that will likely lead to the disruption of a gene product, available in both humans [11, 12, 93] and mice [94], or through inactive dCas9 coupled to either repressors or activators [58, 61].”
BackgroundMethodBogerd, H. P., Kornepati, A. V. R., Marshall, J. B., Kennedy, E. M., & Cullen, B. R. (2015) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1516305112 · Cited by 62
“Although the paper initially describing the SAM methodology reported no evidence of synergy when multiple sgRNAs were used (12), we also wished to ask if we could see detectable synergy between sgRNAs in our system, given previous Fig.”
BackgroundMethodResultInfluentialGerace, D., Martiniello‐Wilks, R., Nassif, N. T., Lal, S., Steptoe, R. J., & Simpson, A. M. (2017) · Stem Cell Research & Therapy · doi:10.1186/s13287-017-0511-8 · Cited by 62
“modified sgRNAs in conjunction with a synergistic activator complex have been shown to induce robust gene activation [46].”
Rapid chromatin repression by Aire provides precise control of immune tolerance
Koh, A. S., Miller, E. L., Buenrostro, J. D., Moskowitz, D. M., Wang, J., Greenleaf, W. J., Chang, H. Y., & Crabtree, G. R. (2018) · Nature Immunology · doi:10.1038/s41590-017-0032-8 · Cited by 63
“In a distinct approach, we employed an orthogonal recruitment strategy using dCas9 and MS2-binding aptamers 36 in the human cortical thymic epithelium cell line 4D6 37 (Supplementary Fig.”
Genome-Scale CRISPRa Screen Identifies Novel Factors for Cellular Reprogramming
Yang, J. P., Rajan, S. S., Friedrich, M., Lan, G., Zou, X., Ponstingl, H., Garyfallos, D. A., Liu, P., Bradley, A., & Metzakopian, E. (2019) · Stem Cell Reports · doi:10.1016/j.stemcr.2019.02.010 · Cited by 63
“, 2018b) using previously established GoF libraries (Kampmann, 2018; Konermann et al., 2015).”
BackgroundMethodCRISPR-Mediated Activation of Endogenous Gene Expression in the Postnatal Heart
Schoger, E., Carroll, K. J., Iyer, L. M., McAnally, J., Tan, W. H., Liu, N., Noack, C., Shomroni, O., Salinas, G., Groß, J., et al. (2019) · Circulation Research · doi:10.1161/circresaha.118.314522 · Cited by 60
Reyes, Á. P., & Lanner, F. (2017) · Development · doi:10.1242/dev.139683 · Cited by 71
“Once fused to epigenetic modifiers, dCas9 can help in the study of chromatin modifications or act as a transcriptional repressor or activator, which might allow its use in gain-of-function experiments (Gilbert et al., 2013; Konermann et al., 2014).”
BackgroundA CRISPR/molecular beacon hybrid system for live-cell genomic imaging
Wu, X., Mao, S., Yang, Y., Rushdi, M. N., Krueger, C. J., & Chen, A. K. (2018) · Nucleic Acids Research · doi:10.1093/nar/gky304 · Cited by 61
“This difference suggests that the stem–loop 2 region is more amendable to modifications than the other two regions, as shown in previous findings (12,39).”
ResultQi, J., Yang, Z., Lin, T., Ma, L., Wang, Y., Zhang, Y., Gao, C.-C., Liu, K.-L., Li, W., Zhao, A., et al. (2021) · Journal of Experimental & Clinical Cancer Research · doi:10.1186/s13046-020-01814-5 · Cited by 61
“PR-Cas9 complex with three SAM components: dCas9– VP64 (Addgene #61425),MS2–p65–HSF1(Addgene, #61426), and sgRNA (Addgene, #89493) as described previously [31].”
MethodHigh‐throughput screens in mammalian cells using the CRISPR‐Cas9 system
Peng, J., Zhou, Y., Zhu, S., & Wei, W. (2015) · FEBS Journal · doi:10.1111/febs.13251 · Cited by 60
Toward the Development of Epigenome Editing-Based Therapeutics: Potentials and Challenges
Ueda, J., Yamazaki, T., & Funakoshi, H. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms24054778 · Cited by 60
“Furthermore, epigenome modification approaches, such as the use of synergistic activation mediators (SAM), CRISPRon, and CRISPRoff (single artificial genes containing multiple EpiEffectors), which simultaneously express multiple types of EpiEffectors, are more effective than those that express a single type of EpiEffe…”
BackgroundEssential RNA-Based Technologies and Their Applications in Plant Functional Genomics
Teotia, S., Singh, D., Tang, X., & Tang, G. (2016) · Trends in biotechnology · doi:10.1016/j.tibtech.2015.12.001 · Cited by 60
“RNAs to manipulate an entire metabolic pathway governed by many proteins/enzymes and guided by different transcriptional controls (activating or repressing) at various steps [106,107].”
BackgroundPark, J., Kim, J., & Bae, S. (2016) · Bioinformatics · doi:10.1093/bioinformatics/btw103 · Cited by 59
“Furthermore, it provides a powerful way to select multiple optimal target sequences from thousands of genes at once for the creation of a genome-wide library.”
BackgroundMammalian Synthetic Biology: Engineering Biological Systems
Black, J. B., Pérez‐Piñera, P., & Gersbach, C. A. (2017) · Annual Review of Biomedical Engineering · doi:10.1146/annurev-bioeng-071516-044649 · Cited by 60
CRISPhieRmix: a hierarchical mixture model for CRISPR pooled screens
Daley, T., Lin, Z., Lin, X., Liu, Y., Wong, W. H., & Qi, L. S. (2018) · Genome biology · doi:10.1186/s13059-018-1538-6 · Cited by 58
“CRISPR interference and CRISPR activation (CRISPRi and CRISPRa, respectively) pooled screens allow for high-throughput functional interrogation of genes through modifications in gene expression [1–3].”
BackgroundCRISPR-cas9: a powerful tool towards precision medicine in cancer treatment
Xing, H., & Meng, L. (2019) · Acta Pharmacologica Sinica · doi:10.1038/s41401-019-0322-9 · Cited by 61
“Genome-wide phenotypic changes can be achieved by transcriptional activation, and high-throughput screening based on GOF has also been introduced to identify genes inducing drug resistance [27, 56].”
BackgroundInfluentialGupta, S., Jain, A., Syed, S. N., Snodgrass, R. G., Pflüger‐Müller, B., Leisegang, M. S., Weigert, A., Brandes, R. P., Ebersberger, I., Brüne, B., et al. (2018) · OncoImmunology · doi:10.1080/2162402x.2018.1494110 · Cited by 59
“To validate that the STAT3/6 binding sites indeed regulate target gene expression, we used CRISPR interference (CRISPRi).(43,44) We prevented the binding of STAT3/STAT6 to their cognate sites at the CCL18 GRR by transfecting the cells with dCas9-KRAB (catalytically inactive Cas9 fused with KRAB repressor domain) and s…”
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“Specifically, we employed a single guide RNA (sgRNA) design containing two MS2 hairpins recognized by the MCP RNA-binding domains (27).”
MethodCRISPR/Cas9 system: a reliable and facile genome editing tool in modern biology
Javaid, D., Ganie, S. Y., Hajam, Y. A., & Reshi, M. S. (2022) · Molecular Biology Reports · doi:10.1007/s11033-022-07880-6 · Cited by 65
CRISPR-Cas9 for medical genetic screens: applications and future perspectives
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“Subsequently, these regions are sequenced and mapped to the pre-existing sgRNA library to read out sgRNA representation in the selected cell subpopulation 16 19 22 ( fi gure 2).”
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LncRNA regulation: New frontiers in epigenetic solutions to drug chemoresistance
Zhou, Y., Sun, W. C., Qin, Z., Guo, S., Kang, Y., Zeng, S., & Yu, L. (2020) · Biochemical Pharmacology · doi:10.1016/j.bcp.2020.114228 · Cited by 58
Vanneste, M., Feddersen, C. R., Varzavand, A., Zhu, E. Y., Foley, T. M., Zhao, L. Q., Holt, K. H., Milhem, M. M., Piper, R. C., Stipp, C. S., et al. (2020) · Frontiers in Oncology · doi:10.3389/fonc.2020.00442 · Cited by 58
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“To systematically compare the relative transactivation potential of different dCas9-based transcriptional activators, we tested five different platforms (Figure 1A). dCas9-VPR (14), dCas9-SAM (13), dCas9-SunTag (12) and dCas9-p300 (16) are widely used dCas9-based transcriptional activators.”
Epigenome editing for targeted DNA (de)methylation: a new perspective in modulating gene expression
Seem, K., Kaur, S., Kumar, S., & Mohapatra, T. (2024) · Critical Reviews in Biochemistry and Molecular Biology · doi:10.1080/10409238.2024.2320659 · Cited by 59
“…promote synergistic activation and recruitment of transcriptional machinery for greater precision/control over gene activation (chavez et al. 2015, Konermann et al. 2015). constitutive expression of dcas9-Suntag scaffold (dcas9-GcN4/sgRNA-scFv-vP64) (tanenbaum et al. 2014) and scFv-p65-HSF1…”
BackgroundRecent advances in therapeutic gene-editing technologies
Liu, D., Cao, D., & Han, R. (2025) · Molecular Therapy · doi:10.1016/j.ymthe.2025.03.026 · Cited by 70
Hu, J. H., Davis, K. M., & Liu, D. R. (2016) · Cell chemical biology · doi:10.1016/j.chembiol.2015.12.009 · Cited by 61
“…in a single experiment has been achieved, enabling large-scale screens that evaluate the effects of disrupting, activating, or repressing each of many different genes (Gilbert et al., 2014; Koike-Yusa et al., 2014; Konermann et al., 2015; Shalem et al., 2014; Wang et al., 2014; Zhou et al., 2014).”
BackgroundCRISPR/Cas9-mediated genome editing in plants
Liu, X., Xie, C., Si, H., & Yang, J. (2017) · Methods · doi:10.1016/j.ymeth.2017.03.009 · Cited by 63
Genome Engineering for Personalized Arthritis Therapeutics
Adkar, S. S., Brunger, J. M., Willard, V. P., Wu, C., Gersbach, C. A., & Guilak, F. (2017) · Trends in Molecular Medicine · doi:10.1016/j.molmed.2017.08.002 · Cited by 57
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CRISPR and personalized Treg therapy: new insights into the treatment of rheumatoid arthritis
Safari, F., Farajnia, S., Aria, M., Zarredar, H., & Nasrolahi, A. (2018) · Immunopharmacology and Immunotoxicology · doi:10.1080/08923973.2018.1437625 · Cited by 58
Transient Retrovirus-Based CRISPR/Cas9 All-in-One Particles for Efficient, Targeted Gene Knockout
Knopp, Y., Geis, F. K., Heckl, D., Horn, S., Neumann, T., Kuehle, J., Meyer, J., Fehse, B., Baum, C., Morgan, M. A., et al. (2018) · Molecular Therapy — Nucleic Acids · doi:10.1016/j.omtn.2018.09.006 · Cited by 63
“adj) to the sgRNA scaffold.(54,55) To facilitate cloning of targeting protospacer sequences into the sgRNA.”
BackgroundMethodIdentification of Antinorovirus Genes in Human Cells Using Genome-Wide CRISPR Activation Screening
Orchard, R. C., Sullender, M. E., Dunlap, B. F., Balce, D. R., Doench, J. G., & Virgin, H. W. (2018) · Journal of Virology · doi:10.1128/jvi.01324-18 · Cited by 56
“HeLa-CD300lf cells were transduced with pLenti-dCas9VP64-BLAST (14).”
BackgroundSynergistic lethality between BRCA1 and H3K9me2 loss reflects satellite derepressionPreprint
Padeken, J., Zeller, P. J., Towbin, B. D., Katic, I., Kalck, V., Methot, S. P., & Gasser, S. M. (2019) · Genes & Development · doi:10.1101/gad.322495.118 · Cited by 62
Determination of local chromatin interactions using a combined CRISPR and peroxidase APEX2 system
Qiu, W., Xu, Z., Zhang, M., Zhang, D., Fan, H., Li, T., Wang, Q., Liu, P., Zhu, Z., Du, D., et al. (2019) · Nucleic Acids Research · doi:10.1093/nar/gkz134 · Cited by 56
“To optimize the CRISPR-based system, we also tested another type of CRISPR system (66) with different promoter and nuclear localization signal (NLS) (Supplementary Figure S9A).”
MethodCRISPR-based gene expression control for synthetic gene circuits
Santos‐Moreno, J., & Schaerli, Y. (2020) · Biochemical Society Transactions · doi:10.1042/bst20200020 · Cited by 59
The NFIB‐ERO1A axis promotes breast cancer metastatic colonization of disseminated tumour cells
Zilli, F., Ramos, P. M., der Maur, P. A., Jehanno, C., Sethi, A., Coissieux, M., Eichlisberger, T., Sauteur, L., Rouchon, A., Bonapace, L., et al. (2021) · EMBO Molecular Medicine · doi:10.15252/emmm.202013162 · Cited by 57
“Using Cas9-Activators with Synergistic Activation Mediators (SAM) (Konermann et al, 2015), we overexpressed Nfib from its endogenous promoter in the nonmetastatic parental HR1 cell line (Fig EV3A).”
BackgroundMethodInfluentialCRISPR Genome Editing Made Easy Through the CHOPCHOP Website
Labun, K., Krause, M., Cleuren, Y. N. T., & Valen, E. (2021) · Current Protocols · doi:10.1002/cpz1.46 · Cited by 57
“Here, position is typically most important (e.g.”
BackgroundThe Role of Recombinant AAV in Precise Genome Editing
Bijlani, S., Pang, K. M., Sivanandam, V., Singh, A., & Chatterjee, S. (2022) · Frontiers in Genome Editing · doi:10.3389/fgeed.2021.799722 · Cited by 63
“, 2013), gene activation (Cheng et al., 2013; Perez-Pinera et al., 2013; Qi et al., 2013; Konermann et al., 2015), gene suppression, (Bikard et al.”
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“One of the most potent currently used CRISPRa tools is the larger SAM system 66 .”
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CRISPR/Cas9 cleavage efficiency regression through boosting algorithms and Markov sequence profiling
Peng, H., Zheng, Y., Blumenstein, M., Tao, D., & Li, J. (2018) · Bioinformatics · doi:10.1093/bioinformatics/bty298 · Cited by 57
“, 2015), gene expressions (Konermann et al., 2015), and clinical trials (Yin et al.”
BackgroundKlompe, S. E., & Sternberg, S. H. (2018) · The CRISPR Journal · doi:10.1089/crispr.2018.0012 · Cited by 57
CRISPR-Based Tools in Immunity
Simeonov, D. R., & Marson, A. (2019) · Annual Review of Immunology · doi:10.1146/annurev-immunol-042718-041522 · Cited by 57
Exploring liver cancer biology through functional genetic screens
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“Moreover, modifications of the CRISPR/Cas system for transcriptional activation of genes will in the future enable identification of gene essentiality and genetic interaction with gain-of-function CRISPR/Cas9 libraries (Gilbert et al., 2014; Konermann et al., 2014).”
MethodMartella, A., Firth, M. A., Taylor, B., Göppert, A., Cuomo, E. M., Roth, R. G., Dickson, A. J., & Fisher, D. I. (2019) · ACS Synthetic Biology · doi:10.1021/acssynbio.8b00527 · Cited by 56
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Systematic discovery of CRISPR-boosted CAR T cell immunotherapies
Datlinger, P., Pankevich, E. V., Arnold, C. D., Pranckevicius, N., Lin, J., Romanovskaia, D., Schaefer, M., Piras, F., Orts, A.-C., Nemc, A., et al. (2025) · Nature · doi:10.1038/s41586-025-09507-9 · Cited by 63
Network analysis of gene essentiality in functional genomics experiments
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“However, for CRISPRa gain of function screen [8, 9], it remains to see whether network-based analysis can bring any predictive power and result enhancement.”
CRISPR/Cas9: From Genome Engineering to Cancer Drug Discovery
Luo, J. (2016) · Trends in cancer · doi:10.1016/j.trecan.2016.05.001 · Cited by 57
“This study found that BRAF inhibition can be bypassed through the over-expression of genes that either re-activate the MAPK pathway through BRAFindependent mechanisms or activate parallel survival pathways independent of the MAPK pathway [24].”
BackgroundMethodSharma, S., & Petsalaki, E. (2018) · International Journal of Molecular Sciences · doi:10.3390/ijms19040933 · Cited by 54
“in a genome-wide CRISPR activation rather than KO screen [46].”
BackgroundPatchable micro/nanodevices interacting with skin
Bai, W., Kuang, T., Chitrakar, C., Yang, R., Li, S., Zhu, D., & Chang, L. (2018) · Biosensors and Bioelectronics · doi:10.1016/j.bios.2018.09.035 · Cited by 56
CRISPR-Cas Activators for Engineering Gene Expression in Higher Eukaryotes
Casas-Mollano, J. A., Zinselmeier, M. H., Erickson, S. E., & Smanski, M. J. (2020) · The CRISPR Journal · doi:10.1089/crispr.2020.0064 · Cited by 58
“SAM is a potent transcriptional activator, consistently outperforming dCas9-VP64.(33) Conversely, the initial iteration of scRNA was up to three times less effective than dCas9-VP64.”
BackgroundMethodInfluentialPérez-García, V., Lea, G., López‐Jiménez, P., Okkenhaug, H., Burton, G. J., Moffett, A., Turco, M. Y., & Hemberger, M. (2021) · eLife · doi:10.7554/elife.63254 · Cited by 56
Metadata
- Authors
- Konermann, S., Brigham, M. D., Trevino, A. E., Joung, J., Abudayyeh, O. O., Barcena, C., Hsu, P. D., Habib, N., Gootenberg, J. S., Nishimasu, H., Nureki, O., & Zhang, F.
- Affiliations
- Broad Institute (US)
- The University of Tokyo (JP)
- Corresponding author
- Zhang, Feng
- ORCID
- Venue
- Nature
- Published
- 2014-12-10
- Volume
- 517
- Issue
- 7536
- Pages
- 583-588
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC4420636
- OpenAlex
- W2127862779
- Cited by
- 2,825
- References
- 52
- Citations by year
- 2026: 154 · 2025: 204 · 2024: 266 · 2023: 236 · 2022: 279 · 2021: 281 · 2020: 301 · 2019: 270 · 2018: 261 · 2017: 229 · 2016: 214 · 2015: 122 · 2014: 3
- FWCI
- 77.58
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- PMC
- Funders
- Japan Society for the Promotion of Science (26291010)
- National Institute of Mental Health (DP1MH100706)
- National Institute of Neurological Disorders and Stroke (R01NS062849, R01NS073124)
- National Center for Chronic Disease Prevention and Health Promotion (DP1‑MH100706)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chromatin Remodeling and Cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- CRISPR/Cas9, transcriptional activation, long intergenic non-coding RNA, gain-of-function screen, BRAF inhibitor resistance, melanoma, drug resistance, gene expression signatures, gene perturbation
- MeSH terms
- Genetic Engineering, Genetic Testing, Humans, Indoles, Melanoma, Sulfonamides, Reproducibility of Results, Transcriptional Activation, Gene Library, Up-Regulation, Genome, Human, Gene Expression Regulation, Neoplastic, DNA, Complementary, Drug Resistance, Neoplasm, RNA, Untranslated, Cell Line, Tumor, Proto-Oncogene Proteins B-raf, Genetic Loci, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems, CRISPR-Associated Proteins
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