Laxmi Uma Maheswar Rao Jakkula Ph.D.,

US Citizen,

**** ********* *****, *****, *****- 75707 (USA).

SUMMARY

Experienced scientist with a background in Invitro Diagnostics tools development in large and small preclinical models and in vitro systems

Extensive management skills in commercial settings

Proficient in molecular biology, Diagnostics assay development, cellular biology, microscopy, histology, bacteriology and virology

Excellent verbal and written skills, strong interpersonal and teamwork skills.

SKILLS

Laboratory Management: Managing people on the research team for Invitro-Diagnostic assay development studies.

This included interviewing and hiring decisions, training and oversight of undergraduate, graduate, postdoctoral fellows and research technicians.

Maintained laboratory notebooks, drafting and finalizing SOPs, facilitated laboratory meetings and journal club.

Ensured research compliance.

Budget development, ordering and cost control.

On-time/ within-budget delivery of research products including managing data collection, collation, analysis and final report writing.

Doctoral Dissertation: Production, Purification, Characterization and Application of Thermostable and Calcium- Independent α-Amylase of Geobacillus thermoleovorans See citation 9-17

Presenting business plan and final reports to management.

Molecular biology: Agarose gel electrophoresis, Southern and Northern blotting, DNA/ RNA extraction and quantification,

Electro Mobility Shift Assay, Far Western, South Western, PCR, RT-PCR, gene silencing by shRNA/or siRNA, cloning, DNA sequencing, cDNA synthesis and gDNA library construction, 5’ and 3’ RACE, Recombinant DNA techniques, Western blotting, RT-qPCR, viral transduction, liposome- mediated transfection, electroporation and lentiCRISPR-Cas9 gene modulations.

Cellular biology: Culturing human lung epithelial cells (A549 cells), isolation and culture of primary colon cells, cell viability assay, cell cycle and differentiation assays, immunofluorescence, flow cytometry, protein extraction purification

Biochemistry and Biophysics: Media designing and process optimization for the production of microbial metabolites; scale-up experience in airlift and stirred tank bioreactors.

Protein expression, purification, characterization, protein-protein, and protein-nucleic acid interaction techniques including native, SDS- PAGE, Western blotting, immune-precipitation, circular dichroism (CD), light scattering (MALS, DLS), HPLC/FPLC, GC, Prep cell, isoelectric focusing, fluorescence spectroscopy, ultra-filtration, ultracentrifugation techniques. Enzyme activity assays, enzyme kinetics (Km, Vmax determinations).

Microscopy: Light microscopy, fluorescent/laser scanning confocal microscopy, and flow cytometry. Histology including immunocytochemistry, HE staining, and immunofluorescence in tissues.

Virology: Amplification and purification of Rift valley fever virus and lentivirus for in-vivo and in-vitro work.

Preclinical (Animal) vaccine testing using pigs, dogs and rodents: Animal restraint, euthanasia, intramuscular or subcutaneous injection, necropsy, intravenous blood sampling, nasal swabs, oral fluid collection.

Computer skills: Microsoft Word, Excel, PowerPoint, Prism Microsoft Office, Adobe Photoshop, Graph Pad, Sigma Plot, Canvas, Alpha Imager, genomic and proteomic analysis tools such as BLAST and FASTA, multiple sequence alignment by CLUSTALW, sequence manipulations using Vector, NTI suite, Snap Gene. Identification of protein sequence by MASCOT.

Language: Written and oral proficiency in English, Hindi, Telugu.

EDUCATION

Ph.D. Microbiology Department of Microbiology, University of Delhi, New Delhi, India

M.S, Applied Microbiology & Biotechnology, Dr. H.S. Gour Vishwavidyalaya Sagar, (MP) India

Bachelor of Science Degree, Microbiology, Botany, Chemistry & Zoology, S.R &B.G.N.R College, Khammam, (AP) India

EXPERIENCE

Pro- Lab Diagnostics, Texas Nov 2021 – Present

COVID-19 testing for the oral fluids, Nasal swabs, blood processing for RT-LAMP assays, RT-RPA-CRISPR-Cas12a, qRT-PCR assay tests.

Virology: Amplification and purification of Rift valley fever virus, viral titer estimation, lentivirus preparation for in-vivo and in-vitro work.

Managing people on the research team for COVID19 testing studies. FDA approval for clinical testing Invitro devises, EUA approved present CoVID-19 testing kit.

Maintained laboratory notebooks, drafting and finalizing SOPs, facilitated laboratory meetings and journal club.

Ensured research compliance.

Budget development, ordering and cost control.

On-time/ within-budget delivery of research products including managing data collection, collation, analysis and final report writing.

Presenting business plan and final reports to management.

Complement Technology May, 2021 – November, 2021

Purification of human blood proteins (C2, factor B, Factor I)

Biochemistry and Biophysics: Media designing and process optimization for the production of microbial metabolites; scale up experience in airlift and stirred tank bioreactors.

Protein C2 purification, IgG antibodies purification, Factor I, Factor B, immune proteins purification from human blood.

Expression of recombinant proteins, characterization, protein-protein interaction studies, and protein-nucleic acid interaction techniques including native, SDS-PAGE, Western blotting, immune-precipitation, circular dichroism (CD), light scattering, fluorescence spectroscopy, ultra- filtration, ultracentrifugation techniques.

Enzyme activity assays, enzyme kinetics (Km, Vmax determinations).

Kansas State University, Manhattan, KS 2014 – 2019

Research Associate

Used molecular biology, cell biology, tissue culture, histology, flow cytometry and imaging techniques. Used viral production (shRNA mediated lentivirus, and lentiCRISPR-Cas9 methodology) /or delivery, gene modulation (lentiCRISPR –Cas9, siRNA, shRNA mediated lentivirus) and animal models.

CRISPR-Cas9 knockout screening and its validation: Rift Valley Fever Virus (RVFV)

CRISPR (clustered regularly interspaced short palindromic repeats) - associated nuclease Cas9 used to genetically modify specific genomic loci.

Used lentiviral delivery of a genomic-scale CRISPR-Cas9 knockout (GeCKO) library targeting 18,080 genes enabling both negative and positive selection in human lung cells (A549 cell line).

Human lung A549 cells were screened for resistance to RVFV infection. Used next generation sequencing to identify candidate genes essential for RVFV replication.

To validate these results, inhibited these gene expression (sgRNAs) by lentiCRISPR-Cas9 system, and created knockout stable cell line. The established methodology was used to knockout for prion protein expression by using lentiCRISPR-Cas9 in deer brain cell line

Kansas University Medical Center, Kansas City, KS 2010- 2013

Postdoc. Fellow

Agarose gel electrophoresis, Southern and Northern blotting, DNA/ RNA extraction and quantification, electro Mobility Shift Assay, Far Western, South Western, PCR, RT-PCR, gene silencing by shRNA/or siRNA, cloning, DNA sequencing

Trained graduate students, post docs and technicians in mouse, pig, dog dissection and other procedures.

Managed research team: oversight of the animal studies for on-time and on-budget delivery.

Microscopy: Light microscopy, fluorescent/laser scanning confocal microscopy, and flow cytometry.

Histology including immunocytochemistry, HE staining, and immunofluorescence in tissues.

MEMBERSHIP IN PROFESSIONAL SOCIETIES

Association of Microbiologists of India (AMI) 2008- present

American Association of Hematology 2013- present

PROFESSIONAL HONORS & AWARDS

Gold medalist and 1st rank in S.R. & B.G.N.R. Degree College throughout B.Sc.

Fast Track Young Scientist- 2008 award by the Department of Science and Technology from Government of India

Young Scientist Award for Industrial Microbiology–2008 award by the Association of Microbiology of India

UBiA elected for Researcher and received 1st position in the researcher competition panel in New University of Lisbon, Faculty of Sciences and Technology, Portugal for 6 months fellowship

CRWAD The best presentation award-2015 (oral presentation)

Green card issued 2015 in EB1A category

PUBLICATIONS

https://scholar.google.com/citations?hl=en&user=j9STRDoAAAAJ&view_op=list_works&sortby=pubdate

NOTE: Due to different conventions in India vs. USA, my initials and name appears two different ways depending upon which country my publication originated, e.g., L.U.M.R. Jakkula in USA and J.L.U.M. Rao in India.

G. D. Boorgula, Laxmi. U M. Jakkula, B. Jung, T. Gumbo, S. Srivastava (2021). Comparison of rifamycins for efficacy against Mycobacterium avium complex and resistance emergence in the hollow fiber model system FRONT. PHARMACOL., 15 April 2021 https://doi.org/10.3389/fphar.2021.645264

I. Ahmed, B. C. Roy, Laxmi Uma Maheswar Rao Jakkula, D. Subramaniam, P. Dandawate, S. Anant, V. Sampath, S. Umar (2020). Infection-induced signals generated at the plasma membrane epigenetically regulate Wnt signaling in vitro and in vivo .JOURNAL OF BIOLOGICAL CHEMISTRY https://www.jbc.org/article/S0021-9258(17)49912-1/fulltext

Alhassan, H. Liu, J. McGill, A. Cerezo, L.U.M.R. Jakkula, A. Nair, E. Winkley, S. Olson, D. Marlow,

Sahni, H. P. Narra, S. Sahni, J. Henningson and R. R. Ganta (2018) Rickettsia rickettsii whole cell antigens offer protection against Rocky Mountain spotted fever in the canine host. INFECTION AND IMMUNITY (ASM) (In Press).

L.U.M.R. Jakkula, H. Liu, T. Von-Ohlen and R.R. Ganta (2016) Sequence Determinants Spanning -35 motif and Spacer Sequences Impacting Ehrlichia chaffeensis Sigma 70-Dependent Promoter Activity of Two p28 Outer Membrane Protein Genes. OXFORD publication DNA RESEARCH 23 (5): 495– 505.

R.K. Raghavan, D.G. Goodin, M.W. Dryden, A. Hroobi, D. Gordon, C. Cheng, A.D. Nair, L.U.M.R. Jakkula, G.A. Hanzlicek, G.A. Anderson, R.R. Ganta (2016) Heterogeneous Associations of Ecological Attributes with Tick-Borne Rickettsial Pathogens in a Periurban Landscape VECTOR- BORNE AND ZOONOTIC DISEASES 16 (9): 569-576.

P. Chandrakesan, B. Roy, L.U.M.R. Jakkula, I. Ahmed, P. Ramamoorthy, O.Tawfik, R. Papineni, C. Houchen, S. Anant, S. Umar (2014). Utility of a Bacterial Infection Model to Study Epithelial- Mesenchymal Transition (EMT), Mesenchymal-Epithelial Transition (MET) or Tumorigenesis ONCOGENE 33(20): 2639–2654.

P. Chandrakesan, J. L. U.M. Rao, I. Ahmed, B. Roy, R. P. Ramanujam, F. G.Mondalek, K. Bradley, S. Anant, S. LeVine, S. Umar (2013). Differential Effect of β-catenin and NF-kB interplay in the Regulation of Cell Proliferation, Inflammation and Tumorigenesis in Response to Bacterial Infection. PLOS ONE https://doi.org/10.1371/journal.pone.0079432

X.Y. Pang, J. Cao, L. Addington, S. Lovell, K. P. Battaile, N. Zhang, J.U.L.M. Rao, E. A. Dennis, A.R. Moise (2012). Structure/Function Relationships of Adipose Phospholipase A2 Containing a Cys-His- His Catalytic Triad. JOURNAL OF BIOLOGICAL CHEMISTRY 287 (2): 35260– 35274.

J.L.U.M. Rao, P. Chattoraj, S. Mohapatra, and I. Biswas (2011). Regulation of Transcription by SMU.1349, a TetR-family Regulator, in Streptococcus mutans. JOURNAL OF BACTERIOLOGY 193 (23): 6605–6613. J.L.U.M. Rao, P. Sudhakar Reddy, R.N. Mishra, D. Gupta, D. Sahal, N. Tuteja, S.K. Sopory, M.K.Reddy (2010). Thermo and pH stable ATP-independent Chaperone Activity of Heat-Inducible HSP70 from Pennisetum glaucum. PLANT SIGNALING AND BEHAVIOR 5:110-121.

J.L.U.M. Rao and T. Satyanarayana (2009). Hyperthermostable, Ca2+-Independent, and High Maltose-Forming α-Amylase Production by an Extreme Thermophile Geobacillus thermoleovorans: Whole Cell Immobilization. APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY 159: 464-477

J.L.U.M. Rao and T. Satyanarayana (2008). Biophysical and Biochemical Characterization of a Hyperthermostable and Ca2+-independent α-Amylase of an Extreme Thermophile Geobacillus thermoleovorans APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY 150: 205–219.

J.L.U.M. Rao and T. Satyanarayana (2007b). Purification and Characterization of a Hyperthermostable and High Maltogenic α-Amylase of an Extreme Thermophile Geobacillus thermoleovorans. APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY 142: 179-93.

J.L.U.M. Rao and T. Satyanarayana (2007a). Improving Production of Hyperthermostable and High Maltose-forming α-Amylase by an Extreme Thermophile Geobacillus thermoleovorans using Response Surface Methodology and its Applications. BIORESOURCE TECHNOLOGY 98: 345–352.

J.L.U.M. Rao and T. Satyanarayana (2004a). Amelioration in Secretion of Hyperthermostable and Ca2+-Independent α-Amylase of Geobacillus thermoleovorans by Some P{olyamines and their Biosynthesis Inhibitor Methylglyoxal-Bis-Guanylhydrazone. JOURNAL OF APPLIED MICROBIOLOGY 97: 1015-1020.

J.L.U.M. Rao and T. Satyanarayana (2004b). Improvement in Secretion of α-Amylase by a Theromophile Geobacillus thermoleovorans. INDIAN JOURNAL OF MICROBIOLOGY 44: 281-284.

T. Satyanarayana, S.M. Noorwez, S. Kumar, J.U.L.M. Rao, M. Ezhilvannan and P. Kaur. (2004). Development of an Ideal Starch Saccharification Process using Amylolytic Enzymes from Thermophiles. BIOCHEMICAL SOCIETY TRANSACTIONS 32: 276-278.

J.L.U.M. Rao and T. Satyanarayana (2003b). Statistical Optimization of a High Maltose-Forming, Hyperthermostable and Ca2+-Independent α-Amylase Production by an Extreme Thermophile Geobacillus thermoleovorans using Response Surface Methodology. JOURNAL OF APPLIED MICROBIOLOGY 95: 712-718.

J.L.U.M. Rao and T. Satyanarayana (2003a). Enhanced Secretion and Low Temperature Stabilization of a Hyperthermostable and Ca2+-Independent α-Amylase of Geobacillus thermoleovorans by Surfactants. LETTERS IN APPLIED MICROBIOLOGY 36: 191-196.

J.L.U.M. Rao, G.D.Y. Boorgula, A.L. Leitão (2011). Fungal enzymes: Present Scenario and Future Perspectives. BENTHAM PUBLISHERS Pp. 3- 23.

T. Satyanarayana, J.U.L.M. Rao and M. Ezhilvannan (2006). α-Amylases In: Enzyme Technology (Eds. Panday A, Webb C, Soccol C. R. & Larroche C), SPRINGER PUBLISHERS, Pp. 189-220.

T. Satyanarayana, DC Sharma, J.L.U.M. Rao, M. Ezhilvannan, M. Bharadwaj, and S. Adhikari (2005).

In Microbial Diversity: Current Perspectives and Potential Applications (Eds. T. Satyanarayana and

B.N. Johri), I.K. INTERNATIONAL PUBLISHERS, Pp. 741-768.

R. Malhotra, S. Narang, J.L.U.M. Rao and T. Satyanaryana (2004). In: Microbiology and Biotechnology for Sustainable Development. (P.C. Jain, Ed), CBS PUBLISHERS, Pp. 261-266.

PRESENTATIONS:

L.U.M.R. Jakkula, et al. (2018) Genome-scale CRISPR-Cas9 Knockout Screening and its Validation: Rift valley Fever Virus, Oral presentation in Phi Zeta Research Day, College of Veterinary Medicine, Kansas State University March 6th 2018 Pp.46

L.U.M.R. Jakkula et al. (2015) Ehrlichia chaffeensis Sigma 70-Dependent Promoter Activity of two Differentially Expressed p28 Outer Membrane Protein Genes, Oral presentation in CRWAD, Chicago, College of Veterinary Medicine, Kansas State University December 6th 2015

W Fiskus, S. Sharma, J. Qi, J.A. Valenta, L. J. Schaub, B. Shah, K. Peth, L.U.M.R. Jakkula, M. Rodriguez, S. G. T. Devaraj, S. Iyer, J.E. Bradner and K.N. Bhalla (2013). BRD4 Antagonist and Histone Deacetylase Inhibitor: a Novel Synergistic Combination against Human Acute Myeloid Leukemia (AML)

cells (55th Annual meeting of ASH) December 7-10 USA.

B. Shah, L.U.M.R. Jakkula, W. Fiskus, S. Sharma, J. Qi, J.A. Valenta, L. J. Schaub, M. Rodriguez, S.G.

T. Devaraj, J.E. Bradner and K.N. Bhalla Co-treatment with BRD4 Antagonist and Histone Deacetylase Inhibitor is Synergistically Lethal against Mantle Cell Lymphoma (MCL) cells. Mol Cancer Ther (2013);12 (11 Suppl):A124.

L.U.M.R. Jakkula, I. Ahmed, B.Roy and S. Umar (2012) Bacterial Infection targets Wnt Signaling to Promote Colonic Crypt Hyperplasia. University of Kansas Cancer center (Cancer research symposium), November 8th 2012, Page number: 18.

A. R. Moise, S. Lovell, N. Zhang, K. P. Battaile, A.Wise, L. Addington, J.L. L.U.M.R. Jakkula, P.

Gao (2012) Structure-Function Studies of Adipose Phospholipase. Kansas, Higuchi Bioscience Centre, February, A1-A6 USA.

L.U.M.R. Jakkula, I. Ahmed, B. Roy and S. Umar (2013) Bacterial Infection Targets Wnt Signaling to Promote Colonic Crypt Hyperplasia DDW conference for Poster Presentation USA.

T. Satyanarayana, J.U.L.M. Rao, J. Vashist and M.R. Rajeswari (2005). Hyperthermostable and Ca2+independent α-amylases of an extreme t h e r m o p h i l e G e o b a c i l l u s thermoleovorans. In: Thermophiles 2005 from evolution to revolution 8th international conference on Thermophiles Research, 18-22 September, 2005 Crowne Plaza Surfers Paradise, Gold Coast, Australia. Pp. 27.

J.L.U.M. Rao and T . Satyanarayana (2005). Purification, Kinetics and Applications of Starch-

Hydrolyzing, Hyperthermostable, Ca -Independent α-Amylase of an Extreme Thermophile Geobacillus thermoleovorans In: Microbial Diversity 2005, International Conference, Microbial Diversity: Current Perspectives and Potential Applications, April 16-18, 2005 New Delhi (India).

J.L.U.M. Rao and T. Satyanarayana (2004). Polyamines and their Biosynthesis Inhibitors on the Production of Hyperthermostable and Ca -Independent α-Amylase by Geobacillus thermoleovorans. In: BioTech 2004, 2 National Conference Biotechnology Society of India, Institute of Genomics and Integrative Biology, October, 13th-15th 2004, New Delhi. Pp.46.

J.L.U.M. Rao and T. Satyanarayana (2002). A High Maltose Forming Ca2+- Independent and Thermostable α-Amylase of an Extreme Thermophile Geobacillus thermoleovorans. In: Proceedings of 43rd Annual Conference of Association of Microbiologists of India, December, 11-13, 2002, Hisar Haryana .Pp106.

J.L.U.M. Rao and T. Satyanarayana (2002). Effect of Surfactants on the Secretion of Calcium Independent Thermostable α-Amylase by Geobacillus thermoleovorans NP 54. In: BIOHORIZON 2002 Fourth National Symposium on Biochemical Engineering and Biotechnology, IIT New Delhi, Pp. BE. 20.

J.L.U.M. Rao and T. Satyanarayana (2001). Effect of Different Surfactants and Polyethyleneglycol on the Production of a Hyperthermostable C a l c i u m -Independent α -Amylase by Geobacillus thermoleovorans. In: Thermophiles 2001, International conference on Biology and Biotechnology of Thermophilic Microbes, December 3- 7, 2001, New Delhi, India Pp.92.

Workshop on Bioinformatics for Cyanobacteriologists (2001) held at National Facility for Marine Cyanobacteria, Bharathidasan University, Tiruchirappalli, India from March 17-28, 2001.

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