C-Brick: A New Standard for Assembly of Biological Parts Using Cpf1.
So far, several DNA assembly standards have been developed, enabling scientists to conveniently share and modify characterized DNA parts. However, a majority of the restriction endonucleases used in these standards bear short recognition sites (e.g., 6 bps in BioBrick standard), which are widely distributed and need to be removed before further construction, causing much inconvenience. Although homing endonucleases, which recognize long DNA sequences, can be used for DNA assembly (e.g., iBrick standard), long scars will be left between parts, limiting their application. Here, we introduce a new DNA assembly standard, namely C-Brick, which employs the newly identified class 2 type V CRISPR-Cas systems protein Cpf1 endonuclease. C-Brick integrates both advantages of long recognition sites and short scars. With C-Brick standard, three chromoprotein cassettes were assembled and further expressed in Escherichia coli, producing colorful pigments. Moreover, C-Brick standard is also partially compatible with the BglBrick and BioBrick standards.
Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences , Shanghai 20032, China.|University of Chinese Academy of Sciences , Shanghai 20032, China.
Shi-Yuan
SY
Li
Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences , Shanghai 20032, China.|University of Chinese Academy of Sciences , Shanghai 20032, China.|Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai , Shanghai 201203, China.|State Key Lab of Genetic Engineering & Center for Synthetic Biology; Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University , Shanghai 200032, China.|Department of Microbiology and Li KaShing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital , Shatin, New Territories, Hong Kong SAR, China.
Guo-Ping
GP
Zhao
Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences , Shanghai 20032, China.|University of Chinese Academy of Sciences , Shanghai 20032, China.
Jin
J
Wang
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D004269:DNA, Bacterial / Q000235:genetics; D004720:Endonucleases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering; D017422:Sequence Analysis, DNA; D058615:Synthetic Biology
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004720:Endonucleases
BioBrick; C-Brick; CRISPR; Cpf1; DNA assembly; synthetic biology