Type-V-F Cas12f proteins, also known as Cas14, have drawn significant interest within the diverse CRISPR-Cas nucleases due to their compact size. This study involves analyzing and comparing Cas14-homology proteins in prokaryotic genomes through mining, sequence comparisons, a phylogenetic analysis, and an array/repeat analysis. In our analysis, we identified and mined a total of 93 Cas14-homology proteins that ranged in size from 344 aa to 843 aa. The majority of the Cas14-homology proteins discovered in this analysis were found within the Firmicutes group, which contained 37 species, representing 42% of all the Cas14-homology proteins identified. In archaea, the DPANN group had the highest number of species containing Cas14-homology proteins, a total of three species. The phylogenetic analysis results demonstrate the division of Cas14-homology proteins into three clades: Cas14-A, Cas14-B, and Cas14-U. Extensive similarity was observed at the C-terminal end (CTD) through a domain comparison of the three clades, suggesting a potentially shared mechanism of action due to the presence of cutting domains in that region. Additionally, a sequence similarity analysis of all the identified Cas14 sequences indicated a low level of similarity (18%) between the protein variants. The analysis of repeats/arrays in the extended nucleotide sequences of the identified Cas14-homology proteins highlighted that 44 out of the total mined proteins possessed CRISPR-associated repeats, with 20 of them being specific to Cas14. Our study contributes to the increased understanding of Cas14 proteins across prokaryotic genomes. These homologous proteins have the potential for future applications in the mining and engineering of Cas14 proteins.

Numan Ullah

Naisu Yang

Zhongxia Guan

Kuilin Xiang

Yali Wang

Mohamed Diaby

Cai Chen

Bo Gao

Chengyi Song

Genes

GENES (ISSN 2073-4425) 是一本经过同行评审的国际开放获取期刊，为与基因、遗传学和基因组学相关的研究提供了一个高级论坛。它发表评论、研究文章、通讯和技术说明。论文的长度没有限制，我们鼓励科学家尽可能详细地发表他们的结果。

GENES (ISSN 2073-4425) is a peer-reviewed international open-access journal that provides a high-level forum for research related to genes, genetics, and genomics. It publishes comments, research articles, communications, and technical notes. There is no restriction on the length of papers, and we encourage scientists to present their results as comprehensively as possible.

《Genes》（ISSN 2073-4425）是一本经过同行评审的国际开放获取期刊，专注于基因、遗传学和基因组学相关研究。期刊为研究者提供了一个高级论坛，发表内容包括评论、研究文章、通讯和技术说明。论文长度没有限制，我们鼓励科学家尽可能详细地分享他们的研究成果。

Comparative Analysis and Phylogenetic Insights of Cas14-Homology Proteins in Bacteria and Archaea

This study involves analyzing and comparing Cas14-homology proteins in prokaryotic genomes through mining, sequence comparisons, a phylogenetic analysis, and an array/repeat analysis, suggesting a potentially shared mechanism of action due to the presence of cutting domains in that region.

Category	Quartile
生物学	3
生物学, 遗传学	3

Comparative Analysis and Phylogenetic Insights of Cas14-Homology Proteins in Bacteria and Archaea

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Journal Info

Category	Quartile
GENETICS & HEREDITY	2