Homo sapiens Raw sequence reads
Engineered T cells expressing chimeric antigen receptors (CARs) or T cell receptors (TCRs) have transformed cancer immunotherapy and are increasingly explored for autoimmune and infectious diseases. Precise genome engineering approaches, including CRISPR-Cas9 mediated gene disruption and targeted integration of large DNA payloads, enable systematic interrogation and enhancement of T cell function. However, efficient and cell type specific delivery of genome editing reagents and donor templates remains a major barrier, particularly for in vivo editing of primary human T cells.To address this challenge, we developed and characterized synthetic adeno-associated virus (AAV) vectors with enhanced tropism for T cells. We evolved HT7, a synthetic AAV with high transduction efficiency in primary human T cells, and performed a genome-wide CRISPR knockout screen to identify host factors required for viral entry and transduction. High throughput sequencing of pooled sgRNA libraries generated raw FASTQ datasets, which were analyzed using MAGeCK to identify genes regulating HT7 mediated transduction, including the identification of CD7 as an essential factor.This dataset includes genome-wide CRISPR screening data, viral transduction assays, and targeted knock-in experiments in human T cells. Raw sequencing reads, processed sgRNA count matrices, and gene level enrichment results are provided to enable reproducible analysis of AAV tropism, host factor dependency, and genetic determinants of efficient T cell transduction. These data support the development of nucleofection free DNA delivery, CRISPR-Cas9 mediated gene editing, and targeted integration of large transgenes in T cells, and provide a resource for studying viral entry mechanisms and optimizing gene delivery strategies for in vivo T cell therapies.