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Wyss Institute for Biologically Inspired Engineering

2 ARTICLES PUBLISHED IN JoVE

JoVE Journal

CRISPR Guide RNA Cloning for Mammalian Systems

Sathiji Nageshwaran^*1,2, Alejandro Chavez^*1,2,3, Nan Cher Yeo^1,2, Xiaoge Guo^1,2, Alissa Lance-Byrne¹, Angela Tung¹, James J. Collins^1,4,5,6,7, George M. Church^1,2

¹Wyss Institute for Biologically Inspired Engineering, Harvard University, ²Department of Genetics, Harvard Medical School, ³Department of Pathology, Massachusetts General Hospital, ⁴Institute for Medical Engineering & Science, Massachusetts Institute of Technology, ⁵Synthetic Biology Center, Massachusetts Institute of Technology, ⁶Department of Biological Engineering, Massachusetts Institute of Technology, ⁷Broad Institute

Here, a simple, efficient, and cost-effective method of sgRNA cloning is outlined.

Bioengineering

Cell-free Protein Expression Using the Rapidly Growing Bacterium Vibrio natriegens

Daniel J. Wiegand^1,2, Henry H. Lee¹, Nili Ostrov^*1, George M. Church^*1,2

¹Department of Genetics, Harvard Medical School, ²Wyss Institute for Biologically Inspired Engineering

Cell-free expression systems are powerful and cost-efficient tools for the high-throughput synthesis and screening of important proteins. Here, we describe the preparation of cell-free protein expression system using Vibrio natriegens for the rapid protein production using plasmid DNA, linear DNA, and mRNA template.