1.Reprogramming Mycobacterium tuberculosis CRISPR System for Gene Editing and Genome-wide RNA Interference Screening
Rahman KHAISTA ; Jamal MUHAMMAD ; Chen XI ; Zhou WEI ; Yang BIN ; Zou YANYAN ; Xu WEIZE ; Lei YINGYING ; Wu CHENGCHAO ; Cao XIAOJIAN ; Tyagi ROHIT ; Naeem Ahsan MUHAMMAD ; Lin DA ; Habib ZESHAN ; Peng NAN ; F.Fu ZHEN ; Cao GANG
Genomics, Proteomics & Bioinformatics 2022;(6):1180-1196
Mycobacterium tuberculosis is the causative agent of tuberculosis(TB),which is still the leading cause of mortality from a single infectious disease worldwide.The development of novel anti-TB drugs and vaccines is severely hampered by the complicated and time-consuming genetic manipulation techniques for M.tuberculosis.Here,we harnessed an endogenous type Ⅲ-A CRISPR/Cas10 system of M.tuberculosis for efficient gene editing and RNA interference(RNAi).This simple and easy method only needs to transform a single mini-CRISPR array plasmid,thus avoiding the introduction of exogenous protein and minimizing proteotoxicity.We demonstrated that M.tuberculosis genes can be efficiently and specifically knocked in/out by this system as con-firmed by DNA high-throughput sequencing.This system was further applied to single-and multiple-gene RNAi.Moreover,we successfully performed genome-wide RNAi screening to identify M.tuberculosis genes regulating in vitro and intracellular growth.This system can be extensively used for exploring the functional genomics of M.tuberculosis and facilitate the development of novel anti-TB drugs and vaccines.
2.The objective assessment of the effects on cognition functioning among military personnel exposed to hypobaric-hypoxia: A pilot fMRI study
Nisha Syed Nasser ; Fathinul Fikri Ahmad Saad ; Aida Abdul Rashid ; Salasiah Mustafa ; Hamed Sharifat ; Rohit Tyagi ; Amei Farina Abd Rashid ; Loh Jia Ling ; Mazlyfarina Mohamad ; Subapriya Suppiah
The Medical Journal of Malaysia 2020;75(1):62-67
Objective: To identify regions of the brain affected during
cognitive working memory during tasks to assess attention,
planning and decision making among military aviation
personnel who have chronic intermittent exposure to high
altitude environment.
Method: A case-control study was conducted in the
Universiti Putra Malaysia among eight military personnel,
four of whom had chronic intermittent exposure to high
altitude training. They were divided into two groups, chronic
intermittent exposure group (CE) (n=4) and a control group
(n=4). They underwent a task-based functional magnetic
resonance imaging (fMRI) that utilised spatial working
memory task to objectively evaluate the neural activation in
response to the Tower of London paradigm. Each correct
answer was given a score of one and the maximum
achievable score was 100%.
Results: A consecutive dichotomised group of CE (4/8) and
control (4/8) of age-matched military aviation personnel with
a mean age of 37.23±5.52 years; showed significant
activation in the right middle frontal gyrus (MFG). This in
turn was positively correlated with response accuracy. A
significant difference in the response accuracy was noted
among both the groups at p<0.05.
Conclusion: At the minimum results of power analysis of
this preliminary fMRI study, our group of aviation personnel
who had chronic intermittent exposure to hypobaric hypoxic
environment, did not have any significant decrease in
cognitive function namely attention, decision-making and
problem solving compared to controls during a working
memory task