A <i>RIPOR2</i> in-frame deletion is a frequent and highly penetrant cause of adult-onset hearing loss

Bruijn, Suzanne E. de; Smits, Jeroen J.; Liu, Chang; Lanting, Cornelis P.; Beynon, Andy J.; Blankevoort, Joëlle; Oostrik, Jaap; Koole, Wouter; Vrieze, Erik de; Cremers, C.W.R.J.; Cremers, Frans P.M.; Roosing, Susanne; Kunst, Henricus P. M.; Zhao, Bo; Pennings, Ronald J.E.; Kremer, Hannie

doi:10.1136/jmedgenet-2020-106863

Cited by 15 publications

(31 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The pathogenicity of this variant has been under debate after it was identified in 2003 in homozygous state in two individuals with normal visual function [16,17]. The second variant is a small in-frame deletion in RIPOR2 (c.1696_1707del; p.(Gln566_Lys569del), NM_014722.3) that we recently identified as a major cause of dominantly inherited hearing loss DFNA21 [18].…”

Section: Introductionmentioning

confidence: 99%

Efficient Generation of Knock-In Zebrafish Models for Inherited Disorders Using CRISPR-Cas9 Ribonucleoprotein Complexes

Vrieze

Bruijn

Reurink

et al. 2021

IJMS

Self Cite

View full text Add to dashboard Cite

CRISPR-Cas9-based genome-editing is a highly efficient and cost-effective method to generate zebrafish loss-of-function alleles. However, introducing patient-specific variants into the zebrafish genome with CRISPR-Cas9 remains challenging. Targeting options can be limited by the predetermined genetic context, and the efficiency of the homology-directed DNA repair pathway is relatively low. Here, we illustrate our efficient approach to develop knock-in zebrafish models using two previously variants associated with hereditary sensory deficits. We employ sgRNA-Cas9 ribonucleoprotein (RNP) complexes that are micro-injected into the first cell of fertilized zebrafish eggs together with an asymmetric, single-stranded DNA template containing the variant of interest. The introduction of knock-in events was confirmed by massive parallel sequencing of genomic DNA extracted from a pool of injected embryos. Simultaneous morpholino-induced blocking of a key component of the non-homologous end joining DNA repair pathway, Ku70, improved the knock-in efficiency for one of the targets. Our use of RNP complexes provides an improved knock-in efficiency as compared to previously published studies. Correct knock-in events were identified in 3–8% of alleles, and 30–45% of injected animals had the target variant in their germline. The detailed technical and procedural insights described here provide a valuable framework for the efficient development of knock-in zebrafish models.

show abstract

Section: Introductionmentioning

confidence: 99%

Efficient Generation of Knock-In Zebrafish Models for Inherited Disorders Using CRISPR-Cas9 Ribonucleoprotein Complexes

Vrieze

Bruijn

Reurink

et al. 2021

IJMS

Self Cite

View full text Add to dashboard Cite

show abstract

“…Structural variants (SVs) were called using the Manta Structural Variant Caller V.1.1.0 (SV detection based on paired end and split read evidence) (Chen et al 2016) and CNVs using Control-FREEC (CNV detection based on alterations in read depth) (Boeva et al 2012). MIP design, sequencing and data analysis were performed as previously described (de Bruijn et al 2021;Neveling et al 2017). MIPs were designed to cover exons and exon-intron boundaries of a panel of 120 HL genes (Online Resource Table S3).…”

Section: Next-generation Sequencing and Variant Interpretationmentioning

confidence: 99%

Exploring the missing heritability in subjects with hearing loss, enlarged vestibular aqueducts, and a single or no pathogenic SLC26A4 variant

et al. 2021

Self Cite

View full text Add to dashboard Cite

Pathogenic variants in SLC26A4 have been associated with autosomal recessive hearing loss (arHL) and a unilateral or bilateral enlarged vestibular aqueduct (EVA). SLC26A4 is the second most frequently mutated gene in arHL. Despite the strong genotype–phenotype correlation, a significant part of cases remains genetically unresolved. In this study, we investigated a cohort of 28 Dutch index cases diagnosed with HL in combination with an EVA but without (M0) or with a single (M1) pathogenic variant in SLC26A4. To explore the missing heritability, we first determined the presence of the previously described EVA-associated haplotype (Caucasian EVA (CEVA)), characterized by 12 single nucleotide variants located upstream of SLC26A4. We found this haplotype and a delimited V1-CEVA haplotype to be significantly enriched in our M1 patient cohort (10/16 cases). The CEVA haplotype was also present in two M0 cases (2/12). Short- and long-read whole genome sequencing and optical genome mapping could not prioritize any of the variants present within the CEVA haplotype as the likely pathogenic defect. Short-read whole-genome sequencing of the six M1 cases without this haplotype and the two M0/CEVA cases only revealed previously overlooked or misinterpreted splice-altering SLC26A4 variants in two cases, who are now genetically explained. No deep-intronic or structural variants were identified in any of the M1 subjects. With this study, we have provided important insights that will pave the way for elucidating the missing heritability in M0 and M1 SLC26A4 cases. For pinpointing the pathogenic effect of the CEVA haplotype, additional analyses are required addressing defect(s) at the RNA, protein, or epigenetic level.

show abstract

“…Several studies have indicated that a loss-offunction mutation in RIPOR2 is correlated with hearing [41][42][43][44]. Diaz-Horta et al identified a splice site mutation (c.102-1G>A) in the RIPOR2 gene (MIM611410) that completely cosegregated with the phenotype in a large consanguineous Turkish family with recessive nonsyndromic, prelingual, profound hearing loss [41].…”

Section: Ripor2 and Hair Cell Functionsmentioning

confidence: 99%

“…Given that RIPOR2 inhibits HDAC6 activity and that HDCA6 destabilizes microtubule structures by promoting α-tubulin deacetylation, it might be possible that RIPOR2 contributes to microtubule flexibility rather than lengthening kinocilia in inner and outer hair cells [43,46]. However, it might be possible that RIPOR2 functions in maintaining normal hearing through mechanisms other than affecting the MYH9 abundance or inhibiting HDAC6 activity [44]. An inframe deletion (c.1696_1707del) mutation in RIPOR2 has been linked with hearing loss [44].…”

Section: Ripor2 and Hair Cell Functionsmentioning

confidence: 99%

Role of RHO family interacting cell polarization regulators (RIPORs) in health and disease: Recent advances and prospects

Lv¹,

Ding²,

Cao³

et al. 2022

Int. J. Biol. Sci.

View full text Add to dashboard Cite

The RHO GTPase family has been suggested to play critical roles in cell growth, migration, and polarization. Regulators and effectors of RHO GTPases have been extensively explored in recent years. However, little attention has been given to RHO family interacting cell polarization regulators (RIPORs), a recently discovered protein family of RHO regulators. RIPOR proteins, namely, RIPOR1-3, bind directly to RHO proteins (A, B and C) via a RHO-binding motif and exert suppressive effects on RHO activity, thereby negatively influencing RHO-regulated cellular functions. In addition, RIPORs are phosphorylated by upstream protein kinases under chemokine stimulation, and this phosphorylation affects not only their subcellular localization but also their interaction with RHO proteins, altering the activation of RHO downstream targets and ultimately impacting cell polarity and migration. In this review, we provide an overview of recent studies on the function of RIPOR proteins in regulating RHO-dependent directional movement in immune responses and other pathophysiological functions.

show abstract

A RIPOR2 in-frame deletion is a frequent and highly penetrant cause of adult-onset hearing loss

Cited by 15 publications

References 58 publications

Efficient Generation of Knock-In Zebrafish Models for Inherited Disorders Using CRISPR-Cas9 Ribonucleoprotein Complexes

Efficient Generation of Knock-In Zebrafish Models for Inherited Disorders Using CRISPR-Cas9 Ribonucleoprotein Complexes

Exploring the missing heritability in subjects with hearing loss, enlarged vestibular aqueducts, and a single or no pathogenic SLC26A4 variant

Role of RHO family interacting cell polarization regulators (RIPORs) in health and disease: Recent advances and prospects

Contact Info

Product

Resources

About