A5058387827- CRISPR and Genetic Engineering
- Advanced biosensing and bioanalysis techniques
- RNA and protein synthesis mechanisms
- RNA regulation and disease
- Pluripotent Stem Cells Research
- Innovation and Socioeconomic Development
- Virus-based gene therapy research
- Photosynthetic Processes and Mechanisms
- RNA Interference and Gene Delivery
- Chromosomal and Genetic Variations
- Plant Virus Research Studies
- Cytomegalovirus and herpesvirus research
- DNA and Nucleic Acid Chemistry
- Retinal Development and Disorders
- Plant tissue culture and regeneration
- Evolution and Genetic Dynamics
- Insect symbiosis and bacterial influences
- RNA modifications and cancer
- Genetics, Aging, and Longevity in Model Organisms
- Viral Infectious Diseases and Gene Expression in Insects
- CAR-T cell therapy research
- Insect Resistance and Genetics
- Viral Infections and Immunology Research
- Mosquito-borne diseases and control
- Skin and Cellular Biology Research
Seoul National University
2009-2025
Seoul National University Hospital
2022-2025
New Generation University College
2023-2025
National University College
2024-2025
Hanyang University
2015-2024
Myongji University
2024
Korea Advanced Institute of Science and Technology
2024
Kangwon National University
2024
Institute of Molecular Biology and Genetics
2024
Anyang University
2021
Abstract Summary: The Type II clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system is an adaptive immune response in prokaryotes, protecting host cells against invading phages or plasmids by cleaving these foreign DNA species a targeted manner. CRISPR/Cas-derived RNA-guided engineered nucleases (RGENs) enable genome editing cultured cells, animals and plants, but are limited off-target mutations. Here, we present novel algorithm termed Cas-OFFinder that searches for...
RNA-guided endonucleases (RGENs), derived from the prokaryotic adaptive immune system known as CRISPR/Cas, enable targeted genome engineering in cells and organisms. RGENs are ribonucleoproteins that consist of guide RNA Cas9, a protein component originated Streptococcus pyogenes . These enzymes cleave chromosomal DNA, whose sequence is complementary, to manner, producing site-specific DNA double-strand breaks (DSBs), repair which gives rise modifications. Despite broad interest...
Abstract Summary: We present Cas-Designer, a user-friendly program to aid researchers in choosing appropriate target sites gene of interest for type II CRISPR/Cas-derived RNA-guided endonucleases, which are now widely used biomedical research and biotechnology. Cas-Designer rapidly provides the list all possible guide RNA sequences given input DNA sequence their potential off-target including bulge-type genome choice. In addition, assigns an out-of-frame score each site help users choose...
Genome editing with programmable nucleases has been widely adopted in research and medicine. Next generation sequencing (NGS) platforms are now used for measuring the frequencies of mutations induced by CRISPR-Cas9 other nucleases. Here, we present an online tool, Cas-Analyzer, a JavaScript-based implementation NGS data analysis. Because Cas-Analyzer is completely at client-side web browser on-the-fly, there no need to upload very large datasets server, time-consuming step genome Currently,...
Abstract Microalgae are versatile organisms capable of converting CO 2 , H O and sunlight into fuel chemicals for domestic industrial consumption. Thus, genetic modifications microalgae enhancing photosynthetic productivity biomass bio-products generation crucial both academic applications. However, targeted mutagenesis in with CRISPR-Cas9 is limited. Here we report, a one-step transformation Chlamydomonas reinhardtii by the DNA-free method rather than plasmids that encode Cas9 guide RNAs....
Abstract Cas12a (also called Cpf1) is a representative type V-A CRISPR effector RNA-guided DNA endonuclease, which provides an alternative to II CRISPR–Cas9 for genome editing. Previous studies have revealed that has unique features distinct from Cas9, but the detailed mechanisms of target searching and cleavage by are still unclear. Here, we directly observe this entire process using single-molecule fluorescence assays study Acidaminococcus sp. (AsCas12a). We determine AsCas12a...
As a result of its simplicity and high efficiency, the CRISPR-Cas system has been widely used as genome editing tool. Recently, CRISPR base editors, which consist deactivated Cas9 (dCas9) or nickase (nCas9) linked with cytidine guanine deaminase, have developed. Base tools will be very useful for gene correction because they can produce highly specific DNA substitutions without introduction any donor DNA, but dedicated web-based to facilitate use such not yet developed.We present two web...
Lutein and zeaxanthin are dietary carotenoids reported to be protective against age-related macular degeneration. Recently, the green alga Chlamydomonas reinhardtii has received attention as a photosynthetic cell factory, but potential of this for carotenoid production not yet been evaluated. In study, we selected C. CC-4349 strain best candidate among seven laboratory strains tested production. A knock-out mutant epoxidase gene induced by preassembled DNA-free CRISPR-Cas9 ribonucleoproteins...
Abstract The type II CRISPR-associated protein Cas9 recognizes and cleaves target DNA with the help of two guide RNAs (gRNAs; tracrRNA crRNA). However, detailed mechanisms kinetics these gRNAs in nuclease activity are unclear. Here, we investigate structural roles CRISPR-Cas9 system by single-molecule spectroscopy reveal a new conformation inactive that is thermodynamically more preferable than active apo-Cas9. We find prevents from changing into form leads to Cas9:gRNA complex. For complex,...
Approximately 15% of non-small cell lung cancer cases are associated with a mutation in the epidermal growth factor receptor (EGFR) gene, which plays critical role tumor progression. With goal treating mutated EGFR-mediated cancer, we demonstrate use clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR protein 9 (Cas9) system to discriminate between oncogenic mutant and wild-type EGFR alleles eliminate carcinogenic allele high accuracy. We targeted an oncogene harboring...
Circulating tumor DNA (ctDNA) has emerged as a tumor-specific biomarker for the early detection of various cancers. To date, several techniques have been devised to enrich extremely small amounts ctDNA present in plasma, but they are still insufficient cancer diagnosis, especially at stage. Here, we developed novel method, CUT (CRISPR-mediated, Ultrasensitive Target DNA)-PCR, which uses CRISPR endonucleases and detect fragments among much more abundant wild-type by specifically eliminating...
Abstract Prime editing technology is capable of generating targeted insertions, deletions, and base conversions. However, the process designing prime guide RNAs (pegRNAs), which contain a primer binding site reverse-transcription template at 3′ end, more complex than that for single used with CRISPR nucleases or editors. Furthermore, assessment high-throughput sequencing data after editors (PEs) have been employed should consider unique feature PEs; thus, pre-existing tools cannot directly...
Prime editing is a versatile and precise genome technique that can directly copy desired genetic modifications into target DNA sites without the need for donor DNA. This holds great promise analysis of gene function, disease modeling, correction pathogenic mutations in clinically relevant cells such as human pluripotent stem (hPSCs). Here, we comprehensively tested prime hPSCs by generating doxycycline-inducible platform. successfully induced all types nucleotide substitutions small...
Cytosine and adenine base editor ribonucleoproteins show precise editing with reduced DNA RNA off-target effects.
Recessive dystrophic epidermolysis bullosa (RDEB) is a severe skin fragility disorder caused by loss-of-function mutations in the COL7A1 gene, which encodes type VII collagen (C7), protein that functions adherence. From 36 Korean RDEB patients, we identified total of 69 pathogenic (40 variants without recurrence), including point (72.5%) and insertion/deletion (27.5%). For fibroblasts from two patients (Pat1 Pat2), applied adenine base editors (ABEs) to correct mutation or bypass premature...
Z-DNA, a left-handed isoform of Watson and Crick’s B-DNA, is rarely formed without the help high salt concentrations or negative supercoiling. However, Z-DNA-binding proteins can efficiently convert specific sequences B conformation into Z in relaxed DNA under physiological conditions. As case many other interactions coupled with structural rearrangements biology, it has been an intriguing question whether actively induce Z-DNAs passively trap transiently preformed Z-DNAs. In this study, we...