2 research outputs found
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 (<i>e.g.</i>, 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 (<i>e.g.</i>, 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 <i>Escherichia coli</i>, producing
colorful pigments. Moreover, C-Brick standard is also partially compatible
with the BglBrick and BioBrick standards
CADS: CRISPR/Cas12a-Assisted DNA Steganography for Securing the Storage and Transfer of DNA-Encoded Information
Because DNA has the merit of high
capacity and complexity, DNA
steganography, which conceals DNA-encoded messages, is very promising
in information storage. The classical DNA steganography method hides
DNA with a “secret message” in a mount of junk DNA,
and the message can be extracted by polymerase chain reaction (PCR)
using specific primers (key), followed by DNA sequencing and sequence
decoding. As leakage of the primer information may result in message
insecurity, new methods are needed to better secure the DNA information.
Here, we develop a pre-key by either mixing specific primers (real
key) with nonspecific primers (fake key) or linking a real key with
3′-end redundant sequences. Then, the single-stranded DNA (ssDNA) <i>trans</i> cleavage activity of CRISPR/Cas12a is employed to
cut a fake key or remove the 3′-end redundant sequences, generating
a real key for further information extraction. Therefore, with the
Cas12a-assisted DNA steganography method, both storage and transfer
of DNA-encoding data can be better protected