The number of fully sequenced genomes increases steadily but the function of many genes remains unstudied. To accelerate dissection of gene function in Leishmania spp. and other kinetoplastids we previously developed a streamlined pipeline for CRISPR-Cas9 gene editing, which we termed LeishGEdit. To facilitate high-throughput mutant screens we have adapted this pipeline by barcoding mutants with unique 17-nucleotide barcodes, allowing loss-of-function screens in mixed populations. Here we present primer design and analysis tools that facilitate these bar-seq strategies. We have developed a standalone easy-to-use pipeline to design CRISPR primers suitable for the LeishGEdit toolbox for any given genome and have generated a list of 14,995 barcodes. Barcodes and oligo sequences are now accessible through our website www.leishgedit.net allowing researchers to pursue bar-seq experiments in all currently available TriTrypDB genomes (release 41). This will streamline CRISPR bar-seq assays in kinetoplastids, enabling pooled mutant screens across the community.

完全测序的基因组数量稳步增加，但许多基因的功能仍未研究。加快利什曼原虫的基因功能解剖spp。和其他动素体，我们之前开发了用于CRISPR-Cas9基因编辑的简化流程，我们称之为LeishGEdit。为了促进高通量突变体筛选，我们通过对具有独特的17个核苷酸条形码的突变体进行条形码编码来适应这一流程，从而允许在混合人群中进行功能丧失的筛选。在这里，我们介绍了有助于这些条码测序策略的引物设计和分析工具。我们已经开发了一个易于使用的独立管道，以针对任何给定的基因组设计适用于LeishGEdit工具箱的CRISPR引物，并生成了14,995条码的列表。现在可以通过我们的网站www.leishgedit.net访问条形码和寡核苷酸序列，使研究人员可以在所有当前可用的TriTrypDB基因组（版本41）中进行条形码序列实验。这将简化动素体中的CRISPR bar-seq分析，