Jim Sawitzke

GeneForge LLC was founded to meet a growing need in both industry and academia for expert, on-demand guidance and advice in genetic construct design. Drawing on over 40 years of hands-on experience in bacterial genetics, molecular biology, recombineering, viral vectors, CRISPR, and more, I offer precise, step-by-step blueprints for building complex genetic constructs. Whether you're troubleshooting a stubborn project or starting from scratch, I work closely with you to find the most effective and reliable path forward—because I know your constructs matter. You can find me at: https://geneforgellc.com

EMBL has a 9 year rule and I am coming to the end of my final contract that cannot be renewed. Also, it has been decided that my facility should merge with the Gene Editing & Embryology Facility. Thus from 1 Jan. 2024 - 14 June 2024 I am a consulting for the new Gene Editing & Virology Facility and easing this transition into a new, combined facility.

The mission of the Genetic & Viral Engineering Facility (GAVEF) is to cater to all viral and genetic engineering needs of EMBL, be it advice, reagents, design, creation, or all of the above. I started this facility after CRISPR reduced the need for complex BAC constructs. In the beginning with one tech, later added a second and now also have a postdoc who is developing new CRISPR-based technologies that can be delivered in a single rAAV and targeted to specific cells. GAVEF specializes in the design and construction of custom DNA vectors and uses them to produce, purify and quantitate viral stocks of rAAV, lentivirus and very recently HSV. In a typical year, we have over 100 such projects. Protocols are optimized for each virus type and new protocols/techniques are routinely tested to optimize yield, purity and precision of quantitation. In addition to custom-made viral vectors and stocks, we provide general sub-cloning services and produce a wide range of DNA targeting constructs for genome engineering (for knock-out, knock-in, conditional knock-out etc) and reagents for CRISPR/Cas9 including long single-strand DNA (up to 6kb). Such constructs are typically made for the Gene Editing and Embryology Facility (GEEF) and they use them for making transgenic mice or cells. We have a large and growing collection of viral vectors, pre-assembled cassettes, and an extensive genetic engineering toolkit available as well as >5000 bacterial glycerols. When time allows I am also developing a new in vivo cloning technology and along with GEEF, we are developing easier ways to make complex genetic constructs in the mouse and in cells as well as optimizing targeting of larger knock-ins.

Responsible for the design, construction and final assembly of complex DNA constructs using classical cloning, recombineering, Gibson assembly, and CRISPR/Cas9 technologies. This included the generation of targeting vectors and other DNA constructs required for transgenesis (including BAC transgenesis), homologous recombination, and nuclease-mediated genomic modification, in the context of mouse mutagenesis. I coordinated my activities with the Head of the Monterotondo Transgenic Core to generate novel animal and cellular models of human disease. Provided support and advice on recombinant DNA experiments and gene targeting to be carried out by researchers at the Unit, and monitor and advise external collaborators of the outstation on DNA recombineering. • organization and supervision of standard laboratory procedures • direction and coordination of laboratory activities with other core facilities and group leaders • monitoring of consumable status and internal budgeting • administration and coordination of facility safety • participation in interviewing, training and supervision of laboratory personnel • organization and updating of common vector map database(s) • preparation of technical training seminars • Aid instruction of external courses that includes aspects of genome engineering • Develop EMBL courses that include genome engineering

As a biologist at the National Cancer Institute in Frederick, MD, USA, I studied the mechanism(s) and optimization of bacteriophage λ Red-mediated recombination, recombineering. Through basic research on the mechanisms of recombineering I was able to improve the efficiency and thus usefulness of the technology. My idea to mutate wobble positions of codons in the donor DNA for recombineering to avoid the bacterial mismatch repair system in a wild type cells led to a patent.

As a postdoctoral fellow, I studied E. coli chromosome segregation with Dr. Stuart Austin. I established that the MukB protein is involved with chromosome chromosome largely through a role in chromosomal condensation. In addition, I studied localization of the SeqA protein and worked on the plasmid P1 segregation system.