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Yeh, K.-L.

Publications and source records attributed to Yeh, K.-L..

2 recordsLinked to original sources

Super-giga and tiny orchid genomes illuminate evolution of Orchidaceae

Orchidaceae (orchids) is commonly known as one of the largest families of seed plants, and grow in an extensive range of habitats worldwide. In the present study, we generated chromosome-level reference genomes for two orchids using a combination of PacBio, Illumina, and Hi-C sequencing, Cypripedium singchii has the largest genome and chromosomes among the sequenced species so far, with a genome size of 43.19 Gb (1C) with ten chromosomes, and Apostasia fujianica has the smallest known genome and chromosomes in Orchidaceae, with a genome size of 340.90 Mb (1C) with 35 chromosomes. We predicted a total of 32,412 and 21,724 protein-coding genes for C. singchii and A. fujianica, respectively. The overall BUSCO score was 85.01% for C. singchii and 91.80% in A. fujianica. Based on protein-coding sequences from 55 conserved single-copy families across 21 plant species, we constructed a high-confidence phylogenetic tree and estimated the divergence times. The high-quality genomes of super-giga and tiny orchids offer key insight for future evolutionary researches.

genomics↗

A robust platform for BaEVRless-lentiviral synthesis and primary natural killer cell transduction

Lentiviral vectors are invaluable tools for genetic modification in human cells for research, biotechnological and clinical applications. However, certain cell types, such as primary human natural killer (NK) cells, present challenges in lentiviral transduction. Overcoming this limitation requires specific pseudotype modifications. BaEVRless-pseudotyped lentivirus (BaEVRless-LV) has shown promise in efficiently transducing human NK cells, B cells, and hematopoietic stem cells (HSCs). BaEVRless, a modified envelope protein derived from Baboon endogenous retrovirus, targets ASCT receptors in human cells. While effective for several immune cell types, BaEVRless-LV production in standard HEK293T cells is challenging. During lentiviral synthesis, BaEVRless protein induces hyper cell fusion, leading to rapid HEK293T cell death and reduced BaEVRless-LV titers. To solve this problem, we used CRISPR genome editing to knockout (KO) the ASCT2 gene in HEK293T cells, thereby abolishing BaEVRless-induced cell fusion. Using the ASCT2-KO cells and an optimized viral production protocol, we efficiently packaged high titers of BaEVRless-LV encoding various transgenes, including turbogfp, chimeric antigen receptor (CAR), and a pooled CRISPR sgRNA library. Our robust BaEVRless-LV synthesis platform is readily adaptable for manufacturing cell therapeutics and enables advanced research techniques such as CRISPR genetic screens in primary NK cells.

immunology↗