Pooled extracellular receptor-ligand interaction screening using CRISPR activation

Chong, Zheng-Shan; Ohnishi, Shuhei; Yusa, Kosuke; Wright, Gavin J.

doi:10.1186/s13059-018-1581-3

Cited by 51 publications

(49 citation statements)

References 57 publications

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“…This could activate the receptor by relieving repression conferred by the NTF or by utilizing the CTF stalk as a fulcrum in a manner akin to class C receptors that use a rigid body to impart activating conformational changes to the 7TM. ADGRB receptors are enriched in the post-synapse and bind secreted C1q-like proteins and an unknown trans-synaptic ligand (possibly the peripheral membrane-associated RTN4R) to regulate synapse formation and maintenance via the thrombospondin-like repeat (TSR) domains (46,142,182). ADGRL is also enriched in the post-synapse and interacts with both FLRT and Teneurin (TEN) single-pass receptors simultaneously to form trans-synaptic links that promote synaptogenesis (78)(79)(80).…”

Section: Resultsmentioning

confidence: 99%

“…While the C1ql proteins and phosphatidylserine are the most studied ligands for ADGRBs, the trans-presented Reticulon 4 Receptor (RTN4R, or Nogo 66 receptor) was recently proposed to be an ADGRB1 ligand that also interacts through the TSRs (142). RTN4R shares many functional features with ADGRB1: both are enriched in neurons and regulate axonal growth, axonal regeneration and synaptic plasticity (143).…”

Section: Trans-cell-presented Proteinsmentioning

confidence: 99%

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Mechanisms of adhesion G protein–coupled receptor activation

Vizurraga

Adhikari

Yeung

et al. 2020

Journal of Biological Chemistry

114

141

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Adhesion G protein coupled receptors (AGPCRs) are a thirty-three-member subfamily of Class B GPCRs that control a wide array of physiological processes and are implicated in disease. AGPCRs uniquely contain large, self-proteolyzing extracellular regions that range from hundreds to thousands of residues in length. AGPCR autoproteolysis occurs within the extracellular GPCR Autoproteolysis-Inducing (GAIN) domain that is proximal to the N-terminus of the G protein-coupling seven transmembrane spanning (7TM) bundle. GAIN domain-mediated self-cleavage is constitutive and produces two-fragment holoreceptors that remain bound at the cell surface. It has been of recent interest to understand how AGPCRs are activated in relation to their two-fragment topologies. Dissociation of the AGPCR fragments stimulates G protein signaling through the action of the tethered-peptide-agonist stalk that is occluded within the GAIN domain in the holoreceptor form. AGPCRs can also signal independently of fragment dissociation, and a few receptors possess GAIN domains incapable of self-proteolysis. This has resulted in complex theories as to how these receptors are activated in vivo, complicating pharmacological advances. Currently there is no existing structure of an activated AGPCR to support any of the theories. Further confounding AGPCR research is that many of the receptors remain orphans and lack identified activating ligands. In this review, we provide a detailed layout of the current theorized modes of AGPCR activation with discussion of potential parallels to mechanisms used by other GPCR classes. We provide a classification means for the ligands that have been identified and discuss how these ligands may activate AGPCRs in physiological-contexts.

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Section: Resultsmentioning

confidence: 99%

Section: Trans-cell-presented Proteinsmentioning

confidence: 99%

Mechanisms of adhesion G protein–coupled receptor activation

Vizurraga

Adhikari

Yeung

et al. 2020

Journal of Biological Chemistry

114

141

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“…7). CRISPR-mediated screens combine high-throughput, single-cell sequencing technologies with genome-wide sgRNA targeting libraries optimized for gene KO (Sanjana et al, 2014;Doench et al, 2016;Morgens et al, 2017;Wang et al, 2018;Liu et al, 2019), activation (Horlbeck et al, 2016;Joung et al, 2017;Chong et al, 2018;Liu et al, 2018b;Sanson et al, 2018), and silencing (Horlbeck et al, 2016;Liu et al, 2017;Sanson et al, 2018) applications. Recent applications of CRISPR-screening have produced new experimental pipelines that permit the unambiguous contribution of risk-associated genes to disease phenotypes (Thyme et al, 2019) and the determination of cellular-lineage and heredity in developmental studies (McKenna et al, 2016;Raj et al, 2018).…”

Section: Crispr Screensmentioning

confidence: 99%

Genetically Engineering the Nervous System with CRISPR-Cas

Sandoval

Elahi

Ploski

2020

eNeuro

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The multitude of neuronal subtypes and extensive interconnectivity of the mammalian brain presents a substantial challenge to those seeking to decipher its functions. While the molecular mechanisms of several neuronal functions remain poorly characterized, advances in next-generation sequencing (NGS) and gene-editing technology have begun to close this gap. The clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (CRISPR-Cas) system has emerged as a powerful genetic tool capable of manipulating the genome of essentially any organism and cell type. This technology has advanced our understanding of complex neurologic diseases by enabling the rapid generation of novel, disease-relevant in vitro and transgenic animal models. In this review, we discuss recent developments in the rapidly accelerating field of CRISPR-mediated genome engineering. We begin with an overview of the canonical function of the CRISPR platform, followed by a functional review of its many adaptations, with an emphasis on its applications for

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“…For the binding screen, the ectodomain of ADGRD1, truncated at S600, was expressed in HEK293-6E cells as a pentameric probe fused to a beta-lactamase enzyme 27 . A Thr to Gly mutation at position 574 in the GPCR proteolytic site (GPS) of human ADGRD1 was introduced to prevent the autoproteolysis 36 .…”

Section: Preparation Of the Protein Library For Large-scale Receptormentioning

confidence: 99%

Control of fluid flow by Adgrd1 is essential for mammalian oviductal embryo transport

Bianchi

Sun²,

Almansa-Ordonez

et al. 2020

Preprint

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Dysfunction of oviductal embryo transport can lead to ectopic pregnancy which affects 1 to 2% of all conceptions in the United States and Europe, and is the most common cause of pregnancy-related death in the first trimester 1, 2 . Ectopic pregnancies almost always occur in the Fallopian tube, emphasizing the critical role of oviductal transport in human reproduction 3 .Oviductal transit is regulated and involves a valve-like "tubal-locking" phenomenon that temporarily arrests oocytes at the ampullary-isthmic junction (AIJ) where fertilization occurs 4 .Here, we show that female mice lacking the orphan adhesion G-protein coupled receptor Adgrd1 are sterile because they are unable to unlock the restraining mechanism at the AIJ, inappropriately retaining embryos within the oviduct. Adgrd1 is expressed on the oviductal epithelium and the post-ovulatory attenuation of tubal fluid production is dysregulated in Adgrd1-deficient mice. We identified Plxdc2 as an activating ligand for Adgrd1 displayed on the surface of cumulus cells. Our findings suggest that regulating oviductal luminal fluid production by Adgrd1 controls embryo transit, and provides important insights into the genetic regulation and molecular mechanisms involved embryo tubal transport.

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Pooled extracellular receptor-ligand interaction screening using CRISPR activation

Cited by 51 publications

References 57 publications

Mechanisms of adhesion G protein–coupled receptor activation

Mechanisms of adhesion G protein–coupled receptor activation

Genetically Engineering the Nervous System with CRISPR-Cas

Control of fluid flow by Adgrd1 is essential for mammalian oviductal embryo transport

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