A5025332400- Telomeres, Telomerase, and Senescence
- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- RNA Research and Splicing
- RNA regulation and disease
- DNA Repair Mechanisms
- RNA modifications and cancer
- Genetics, Aging, and Longevity in Model Organisms
- Advanced biosensing and bioanalysis techniques
- RNA Interference and Gene Delivery
- Amyotrophic Lateral Sclerosis Research
- Fungal and yeast genetics research
- bioluminescence and chemiluminescence research
- Neurobiology and Insect Physiology Research
- Plant Disease Resistance and Genetics
- Cancer-related molecular mechanisms research
- Plant and Fungal Interactions Research
- Neurogenetic and Muscular Disorders Research
- Genomics and Chromatin Dynamics
- Parkinson's Disease Mechanisms and Treatments
University of Colorado Boulder
2019-2024
University of Massachusetts Chan Medical School
2020-2024
Johns Hopkins University
2015-2020
Significance Telomerase reverse transcriptase (hTERT) is involved in immortalization and survival of cancer cells. Recurring mutations its promoter often lead to reexpression cancer. Therapies targeting hTERT activity have been challenging develop, none are routine use. Using targeted functional genomics knockout screening a human bladder model, we found that transcription factor TRIM28 activates expression preferentially from the mutant allele. We also revealed therapeutically targetable...
Abstract The spatial partitioning of the transcriptome in cell is an important form gene-expression regulation. Here, we address how intron retention influences spatio-temporal dynamics transcripts from two clinically relevant genes: TERT (Telomerase Reverse Transcriptase) pre-mRNA and TUG1 (Taurine-Upregulated Gene 1) lncRNA. Single molecule RNA FISH reveals that nuclear uniformly robustly retain specific introns. Our data suggest splicing retained introns occurs during mitosis. In...
In Saccharomyces cerevisiae and in humans, the telomerase RNA subunit is bound by Ku, a ring-shaped protein heterodimer best known for its function DNA repair. Ku binding to yeast promotes telomere lengthening recruitment telomeres, but how this achieved remains unknown. Using telomere-length analysis chromatin immunoprecipitation, we show that Sir4 – previously identified Ku-binding component of telomeric silent required Ku-mediated recruitment. We also find specifically tethering directly...
Telomerase is pathologically reactivated in most human cancers, where it maintains chromosomal telomeres and allows immortalization. Because telomerase reverse transcriptase (TERT) usually the limiting component for activation, numerous studies have measured TERT mRNA levels populations of cells or tissues. In comparison, little known about expression at single-cell single-molecule level. To address this, we analyzed across 10 cancer lines using RNA fluorescent situ hybridization (FISH) made...
Abstract Many of the biological functions performed by RNA are mediated RNA-binding proteins (RBPs), and understanding molecular basis these interactions is fundamental to biology. Here, we present massively parallel assay combined with immunoprecipitation (MPRNA-IP) for in vivo high-throughput dissection RNA–protein describe statistical models identifying domains parsing structural contributions RNA. By using custom pools tens thousands sequences containing systematically designed...
Telomerase RNA contains a template for synthesizing telomeric DNA and has been proposed to act as flexible scaffold holoenzyme protein subunits in the RNP. In Saccharomyces cerevisiae, telomerase RNA, TLC1, is bound by Sm7 complex, which required stabilization of predominant, non-polyadenylated (poly(A)-) TLC1 isoform. However, it remains unclear (1) whether retains this function when its binding site repositioned within shown other TLC1-binding subunits, (2) how stabilizes poly(A)- TLC1....
The telomerase ribonucleoprotein (RNP) counters the chromosome end replication problem, completing genome to prevent cellular senescence in yeast, humans, and most other eukaryotes. RNP core enzyme is composed of a dedicated RNA subunit reverse transcriptase (telomerase [TERT]). Although majority 1,157-nucleotide (nt) Saccharomyces cerevisiae RNA, TLC1, rapidly evolving, central catalytic largely conserved, containing template, template-boundary helix, pseudoknot, core-enclosing helix (CEH)....
GGGGCC (G4C2) repeat expansion in C9ORF72 is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). How this mutation leads to neurodegeneration remains largely unknown. Using CRISPR-Cas9 technology, we deleted EXOC2, which encodes an essential exocyst subunit, induced pluripotent stem cells (iPSCs) derived from C9ORF72-ALS/FTD patients. These are viable owing presence truncated suggesting that function partially maintained. Several...
ABSTRACT Despite the great importance of RNA-protein interactions in cells, there is a very limited set approaches available for identifying proteins that bind to specific RNA. We report here combining use CRISPR technology with yeast two-hybrid protein-protein interaction system order create an advantageous method investigating interactions. In this C RISPR- ssisted R NA/ BP y east (CARRY) system, RNA interest targeted promoters standard reporter genes by fusing it guide strain expressing...
ABSTRACT Telomerase RNA contains a template for synthesizing telomeric DNA by reverse transcription and has been proposed to act as flexible scaffold holoenzyme protein subunits in the RNP. In Saccharomyces cerevisiae , telomerase Est1 Ku bind RNA, TLC1, it shown that these proteins still function when their binding sites are repositioned within RNA. TLC1 is also bound Sm 7 complex, which required stabilization of predominant, non-polyadenylated (poly(A)–) isoform. Here, we first show can...
Abstract Telomerase is pathologically reactivated in most human cancers, where it maintains chromosomal telomeres and allows immortalization. Because telomerase reverse transcriptase (TERT) usually the limiting component for activation, numerous studies have measured TERT mRNA levels populations of cells or tissues. However, little known about expression at single-cell single-molecule level. Here we analyzed across 10 cancer lines using RNA FISH made several unexpected findings. First, there...
ABSTRACT The telomerase RNP counters the chromosome end-replication problem, completing genome replication to prevent cellular senescence in yeast, humans, and most other eukaryotes. core enzyme is composed of a dedicated RNA subunit reverse transcriptase (TERT). Although majority 1157-nt Saccharomyces cerevisiae RNA, TLC1, rapidly evolving, central catalytic largely conserved, containing template, template-boundary helix, pseudoknot, core-enclosing helix (CEH). Here, we show that 4-base...
ABSTRACT Many of the biological functions performed by RNA are mediated RNA-binding proteins (RBPs), and understanding molecular basis these interactions is fundamental to biology. Here, we present MPRNA-immunoprecipitation (MPRNA-IP), an adaptation previously developed massively parallel assay (MPRNA), a new avenue for in vivo high-throughput dissection RNA-protein interactions. By using custom pools tens thousands sequences containing systematically designed truncations mutations, able...