Journal: bioRxiv

Article Title: Multiplex base editing to convert TAG into TAA codons in the human genome

doi: 10.1101/2021.07.13.452007

Figure Lengend Snippet: a, Framework for converting TAG codons into TAA in human cells. b, UAGs number and editable UAG sites of all genes and essential genes in each chromosome. c-e, distribution analysis of cells with different number of modified gene targets in populations with different delivery methods based on single cell RNAseq. Method_1, delivery 10 gBlocks with mCherry-inactivated eGFP reporter; Method_2, delivery 10 gBlocks with mCherry-inactivated eGFP reporter and eGFP sgRNA plasmids; Method_3, delivery 43-all-in-one with DsRed. f, Density plot for distribution of number of modified gene targets detected by scRNAseq in 3 populations. Vertical lines indicate the median values of modified gene targets. g, For each gene target, distribution analysis of modified cells with different editing efficiency. Counts from different methods were stacked in the plot. h, Editing efficiency of each sgRNAs in single cells. i, Heat map of target “C” editing efficiency in cell population with three delivery methods based on converting single-cell RNA-Seq into Bulk RNA-Seq. Editing efficiency was indicated with the intensity of red.

Article Snippet: All gBlocks fragment containing 5 sgRNA expression cassettes with high fidelity four-base overhang pair after cutting with type IIS restriction enzyme BbsI restriction enzyme were designed and directly sent to be synthesized into PUC57 cloning plasmid by GenScript.

Techniques: Modification, RNA Sequencing

Journal: Immunity

Article Title: Global analysis of shared T cell specificities in human non-small cell lung cancer enables HLA inference and antigen discovery

doi: 10.1016/j.immuni.2021.02.014

Figure Lengend Snippet:

Article Snippet: Briefly, synthetic gene blocks (gBlocks®) of N-terminal truncated TCRα or TCRβ chain V and modified C gene fragments were assembled into the baculoviral pAcGP67a construct (BD Biosciences) with Gibson assembly (New England BioLabs).

Techniques: Recombinant, Clone Assay, Plasmid Preparation, Expressing, Construct, Software