Project description:A rare mutation associated with familial ST-wave depression in ECG was predicted to generate a de novo enhancer. This mutation was generated heterozygously in iPSC lines to characterise it's molecular effects by ATAC-seq, Capture-C and RNA-seq
Project description:A rare mutation associated with familial ST-wave depression in ECG was predicted to generate a de novo enhancer. This mutation was generated heterozygously in iPSC lines to characterise it's molecular effects by ATAC-seq, Capture-C and RNA-seq
Project description:SpCas9 INDEL frequency and HITI was evaluated at the on-target site of guide RNA targeting either the mouse or pig rhodopsin by next-generation sequencing.
Project description:SpCas9 INDEL frequency was evaluated at both the on- and off-target sites of guideRNA targeting either the murine Rhodopsin or Albumin by Next Generation Sequencing. Minimal INDELs were found at the off target sites evaluted, while SpCas9 activity was predominantly on-target.
Project description:Across the cell cycle, mitochondrial dynamics are regulated by a cycling wave of actin polymerization/depolymerization. In metaphase, this wave induces the assembly of actin comet tails on mitochondria that propel these organelles to drive spatial mixing,resulting in their equitable inheritance by daughter cells. In contrast, during interphasethe cycling actin wave promotes localized mitochondrial fission. Here, we identify the F-actin nucleator/elongator FMNL1 as a positive regulator of the wave. Depletion of FMNL1 ablates the actin wave, allowing us to assess the functional consequences in interphase cells. FMNL1-depleted cells exhibit decreased mitochondrial polarization, decreased mitochondrial oxygen consumption, and increased production of reactive oxygen species. Accompanying these changes is a loss of hetero-fusion of wave-fragmented mitochondria. Thus we propose that the interphase actin wave maintains mitochondrial homeostasis by promoting mitochondrial content mixing. Finally, we investigated the mechanistic basis for the observation that the wave drives mitochondrial motility in metaphase but mitochondrial fission in interphase. Our data indicate that when the force of actin polymerization is resisted by mitochondrial tethering to microtubules, fission results. In striking contrast, upon microtubule depolymerization actin wave-enveloped mitochondria in interphase cells display comet tail motility characteristic of metaphase cells, which are devoid of microtubule-mitochondria interactions, suggesting that microtubule tethering inhibits comet tail-driven motility.
Project description:Amino acid insertions and deletions (indels) are an abundant class of genetic variants. However, compared to substitutions, the effects of indels are not well understood and poorly predicted. Here we address this shortcoming by performing deep indel mutagenesis (DIM) of structurally diverse proteins. Indel tolerance is strikingly different to substitution tolerance and varies extensively both between different proteins and within different regions of the same protein. Although state of the art variant effect predictors perform poorly on indels, we show that both experimentally-measured and computationally-predicted substitution scores can be repurposed as good indel variant effect predictors by incorporating information on protein secondary structures. Quantifying the effects of indels on protein-protein interactions reveals that insertions can be an important class of gain-of-function variants. Our results provide an overview of the impact of indels on proteins and a method to predict their effects genome-wide.