Dr. Qi Lei Releases New Achievements in CRISPRi Research
Researchers from Stanford University and the University of California, San Francisco reported that they used CRISPR Interference (CRISPRi) technology to perform a comprehensive functional analysis of the essential genes of Bacillus subtilis. The study was published in the May 26 issue of Cell.
Dr. Lei S. Qi of Stanford University, Dr. Kerwyn Casey Huang, and Dr. Carol A. Gross of the University of California, San Francisco are co-authors of the paper. Qi Lei graduated from Tsinghua University in the early years. His current research interests are the use of genomic engineering and CRISPR technology to study genetic interaction networks related to cell differentiation, proliferation, epigenetic regulation and disease. A number of CRISPR articles have been published in the journals Cell, Nature Biotechnology, and Nature Methods.
In 2013, Dr. Qi Lei published a paper in Cell to introduce the gene silencing technology developed by the research team. Qi Lei led researchers to use CRISPR for a platform for gene expression sequence-specific regulation and developed a Cas9-based gene regulation method. They refer to this system as CRISPRi and point out that this RNA-directed DNA recognition platform is a simple new method for selective inhibition of gene expression within the genome.
In January 2015, Qi Lei collaborated with Wendell A. Lim of the University of California, San Francisco to develop a CRISPR scaffold RNA (scRNA). The scRNA encodes a target site and a regulatory function that simultaneously activates and inhibits different genes. The study was published in the journal Cell. This research provides a powerful tool for researchers designing gene expression programs.
In 2015, Dr. Qi Lei collaborated with the Wang Xiaoyi team of the Department of Automation of Tsinghua University to develop a comprehensive calculation tool based on a set of sgRNA design rules summarized in published research. They reported a genome-wide sgRNA design tool and provided an online website for predicting efficient and specific sgRNA. They named the tool CRISPR-ERA for CRISPR-mediated gene editing, suppression and activation.
In this Cell article, Qi Lei and co-authors point out that essential genes function to support the core cellular processes. Investigating the relationship between essential gene functions is critical to understanding the mechanisms that control bacterial growth and driving drug development. However, there are currently only a few methods available to assess essential gene functions in vivo to elucidate their relationship. Since the essential gene loss function can lead to the inability of cells to survive, neither gene deletion libraries nor saturated transposon mutagenesis can be used to study essential genes. Several high-throughput methods have been used to identify or disrupt essential genes in eukaryotes, including instability of mRNAs 3'UTR, CRISPR/Cas9 gene editing, and CRISPR/dCas9 transcriptional regulation techniques. The only current study to screen for essential genes in bacteria uses antisense RNA knockdown to screen for antibiotic susceptibility, which is limited by the effectiveness of this method.
In the new study, researchers used CRISPRi to knock down every essential gene in Bacillus subtilis and explore their phenotype. Their use of high-reliability essential gene networks established by chemical genomics shows a broad interrelationship between distant processes and identifies patterns of action for antibiotics with unidentified characteristics. Importantly, the researchers found that a slight knockdown of essential gene function significantly reduced quiescent survival without affecting the highest growth rate. Finally, high-throughput microscopy analysis showed that cell morphology was relatively insensitive to mild knockdown and was significantly affected by the function of the deleted gene, revealing a close relationship between cell growth and morphology.
The new study provides a framework for a wide range of microbial and comparative analyses for in vivo systematic investigation of essential gene functions.
Hospital Nitrile Gloves,Exam Nitrile Gloves,Nitrile Gloves Bulk,Medical Hand Gloves Powder Free
Puyang Linshi Medical Supplies Co., Ltd. , https://www.linshimedicals.com