Difference between revisions of "Part:BBa K2019000"

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CRISPR-Cas9 has one of the most promising tools for genome editing in the field of synthetic biology due its ability to cut DNA with extremely low off target effects and high efficiency.  The most commonly used CRISPR endonuclease is Cas9 from the bacteria streptococcus pyogenes (spCas9).  However, one major limitation of saCas9 is its size.  The gene that encodes for WT spCas9 is roughly 4.2 kb and produces a protein which is roughly 155 kDa, which runs into issues for several delivery mechanisms, including adeno-associated viruses (AAV) and outer membrane vesicles (our own delivery mechanism).  saCas9 helps alleviate the issue of size of Cas9 with retaining the same functionality as spCas9.  saCas9's gene is only 3.2kb, andd the protein size is around 113 kDa, over 40kDa less than spCas9.  Furthermore, the PAM sequence for saCas9 is NNGRRT, compared to the NGG PAM sequence for spCas9.  Statistically, the increased complexity of the PAM sequence of saCas9 will lead to less off-target effects compared to its counterpart spCas9.

CRISPR-Cas9 has one of the most promising tools for genome editing in the field of synthetic biology due its ability to cut DNA with extremely low off target effects and high efficiency.  The most commonly used CRISPR endonuclease is Cas9 from the bacteria streptococcus pyogenes (spCas9).  However, one major limitation of saCas9 is its size.  The gene that encodes for WT spCas9 is roughly 4.2 kb and produces a protein which is roughly 155 kDa, which runs into issues for several delivery mechanisms, including adeno-associated viruses (AAV) and outer membrane vesicles (our own delivery mechanism).  saCas9 helps alleviate the issue of size of Cas9 with retaining the same functionality as spCas9.  saCas9's gene is only 3.2kb, andd the protein size is around 113 kDa, over 40kDa less than spCas9.  Furthermore, the PAM sequence for saCas9 is NNGRRT, compared to the NGG PAM sequence for spCas9.  Statistically, the increased complexity of the PAM sequence of saCas9 will lead to less off-target effects compared to its counterpart spCas9.