Biosafety Level – 3 (BSL-3) Laboratory

ABOUT

The Biosafety level three facility (BSL-3) is a containment facility located in the Centre for Infectious Disease Research (CIDR), IISc Bangalore for carrying out associated research activities involving airborne pathogens such as Mycobacterium Tuberculosis and other human pathogens including HIV, Hepatitis, Japanese Encephalitis. CIDR BSL-3 facilities have been certified by the Department of Biotechnology, Government of India. The laboratory has special safety and engineering features for maintaining requisite negative pressure environment to ensure unidirectional airflow, and for ensuring the safety of lab personnel as per the CDC and WHO guidelines. A dedicated building management system (BMS) is in place to enable 24×7 monitoring of the critical parameters such as pressure, temperature, and humidity.

OBJECTIVE

• To undertake research on highly pathogenic and infectious disease such as Mtb and other BSL-3 class pathogens.
• The facility will strengthen the research activity in the ongoing anti Mtb and molecular targets projects of Mycobacterium Tuberculosis.
• Apart from the inhouse research activities, this facility will also cater as regional center for providing diagnostics in case of outbreaks.

FACILITIES AVAILABLE

BSL-3

• 4560 sq. feet equipped containment facility with separate sections for BSL-3 and ABSL-3.
• The core BSL-3 laboratory consists of three in-vitro experimentation labs and one infection-cum-necropsy room.
• The remaining area includes service areas such as shower entry area, separate male and female lock rooms, media preparation room, entry corridors, exit corridors, and effluent treatment areas.
• The facility is fully equipped with certified biosafety cabinets, horizontal autoclaves, high speed centrifuges, BD FACS Aria sorter, Seahorse XF flux analyser, multimode plate reader, spectrophotometer, Electroporator, cell culture accessories including CO2 incubators, shaker incubators etc.

ABSL-3

• The ABSL-3 laboratory has an overall footprint of 260 sq. feet.
• The ABSL-3 has IVC system for safe and hygienic housing of small animals (Mice and Guinea pig). The facility houses an animal change station, Wisconsin-Madison aerosol chamber for aerosol route infection of animals, and GentleMACS animal tissue dissociator.
• Various animal studies such as protective efficacy studies of vaccine candidates, therapeutics, evaluation of drug candidates, virulence studies of various Mycobacterium Strains and survival studies can be performed.

SERVICES PROVIDED

• Invitro experiments with Mycobacterium tuberculosis.
• FACS sorter, BD FACS ARIA Fusion.
• Ex-vivo experiments with immune cells (BMDM, Peritoneal Macrophages) etc.
• In vivo animal efficacy/infection models with Mice and Guinea Pigs.

RECENT PUBLICATIONS

1. Shee S, Veetil RT, Mohanraj K, Das M, Malhotra N, Bandopadhyay D, Beig H, Birua S, Niphadkar S, Nagarajan SN, Sinha VK, Thakur C, Rajmani RS, Chandra N, Laxman S, Singh M, Samal A, Seshasayee AN, Singh A. Biosensor-integrated transposon mutagenesis reveals rv0158 as a coordinator of redox homeostasis in Mycobacterium tuberculosis. Elife. 2023 Aug 29;12:e80218. doi: 10.7554/eLife.80218. PMID: 37642294; PMCID: PMC10501769.
2. Borbora SM, Satish BA, Sundar S, B M, Bhatt S, Balaji KN. Mycobacterium tuberculosis elevates SLIT2 expression within the host and contributes to oxidative stress responses during infection. J Infect Dis. 2023 May 9:jiad126. doi: 10.1093/infdis/jiad126. Epub ahead of print. PMID: 37158474.

2. Rakshit S, Adiga V, Ahmed A, Parthiban C, Chetan Kumar N, Dwarkanath P, Shivalingaiah S, Rao S, D’Souza G, Dias M, Maguire TJA, Doores KJ, Zoodsma M, Geckin B, Dasgupta P, Babji S, van Meijgaarden KE, Joosten SA, Ottenhoff THM, Li Y, Netea MG, Stuart KD, De Rosa SC, McElrath MJ, Vyakarnam A. Evidence for the heterologous benefits of prior BCG vaccination on COVISHIELD™ vaccine-induced immune responses in SARS-CoV-2 seronegative young Indian adults. Front Immunol. 2022 Oct 4;13:985938. doi: 10.3389/fimmu.2022.985938. PMID: 36268023; PMCID: PMC9577398.

3. Banerjee U, Rao P, Reddy M, Hussain M, Chunchanur S, Ambica R, Singh A, Chandra N. A 9-gene biomarker panel identifies bacterial coinfections in culture-negative COVID-19 cases. Mol Omics. 2022 Sep 26;18(8):814-820. doi: 10.1039/d2mo00100d. PMID: 35971789.

4. Shee S, Singh S, Tripathi A, Thakur C, Kumar T A, Das M, Yadav V, Kohli S, Rajmani RS, Chandra N, Chakrapani H, Drlica K, Singh A. Moxifloxacin-Mediated Killing of Mycobacterium tuberculosis Involves Respiratory Downshift, Reductive Stress, and Accumulation of Reactive Oxygen Species. Antimicrob Agents Chemother. 2022 Sep 20;66(9):e0059222. doi: 10.1128/aac.00592-22. Epub 2022 Aug 17. PMID: 35975988; PMCID: PMC9487606.

5. Borbora SM, Rajmani RS, Balaji KN. PRMT5 epigenetically regulates the E3 ubiquitin ligase ITCH to influence lipid accumulation during mycobacterial infection. PLoS Pathog. 2022 Jun 3;18(6):e1010095. doi: 10.1371/journal.ppat.1010095. PMID: 35658060; PMCID: PMC9200362.

6. Bandyopadhyay P, Pramanick I, Biswas R, Ps S, Sreedharan S, Singh S, Rajmani RS, Laxman S, Dutta S, Singh A. S-Adenosylmethionine-responsive cystathionine β-synthase modulates sulfur metabolism and redox balance in Mycobacteriumtuberculosis. Sci Adv. 2022 Jun 24;8(25):eabo0097. doi: 10.1126/sciadv.abo0097. Epub 2022 Jun 24. PMID: 35749503; PMCID: PMC9232105.

7. Tripathi A, Anand K, Das M, O’Niel RA, P S S, Thakur C, R L RR, Rajmani RS, Chandra N, Laxman S, Singh A. Mycobacterium tuberculosis requires SufT for Fe-S cluster maturation, metabolism, and survival in vivo. PLoS Pathog. 2022 Apr 15;18(4):e1010475. doi: 10.1371/journal.ppat.1010475. PMID: 35427399; PMCID: PMC9045647.

8. Thakur C, Tripathi A, Ravichandran S, Shivananjaiah A, Chakraborty A, Varadappa S, Chikkavenkatappa N, Nagarajan D, Lakshminarasimhaiah S, Singh A, Chandra N. A new blood-based RNA signature (R₉), for monitoring effectiveness of tuberculosis treatment in a South Indian longitudinal cohort. iScience. 2022 Jan 10;25(2):103745. doi: 10.1016/j.isci.2022.103745. PMID: 35118358; PMCID: PMC8800112.

9. Sharma PR, Dravid AA, Kalapala YC, Gupta VK, Jeyasankar S, Goswami A, Agarwal R. Cationic inhalable particles for enhanced drug delivery to M. tuberculosis infected macrophages. Biomater Adv. 2022 Feb;133:112612. doi: 10.1016/j.msec.2021.112612. Epub 2021 Dec 22. PMID: 35527151.

10. Anand C, Santoshi M, Singh PR, Nagaraja V. Rv0802c is an acyltransferase that succinylates and acetylates Mycobacterium tuberculosisnucleoid-associated protein HU. Microbiology (Reading). 2021 Jul;167(7). doi: 10.1099/mic.0.001058. PMID: 34224344.

11. Fernandes SE, Alakesh A, Rajmani RS, Jhunjhunwala S, Saini DK. Aging associated altered response to intracellular bacterial infections and its implication on the host. Biochim Biophys Acta Mol Cell Res. 2021 Aug;1868(9):119063. doi: 10.1016/j.bbamcr.2021.119063. Epub 2021 May 20. PMID: 34022281.

USER INSTITUES

• IISc, Bangalore
• NCBS, Bangalore
• JNCASR, Bangalore
• NIMHANS, Bangalore
• FNDR, Bangalore
• CDRI, Lucknow
• IISER, Pune
• IISER, Tirupati
• IIT, Roorkee