CRISPR and Viral Diagnostics Susan Eiben Jun Huang Kira Slepchenko httpswwwyoutubecomwatchv22F85FOAyik CRISPRCas9 httpslabiotecheufeaturescrisprcas9reviewgeneeditingtool Clusters of Regularly Interspaced Short Palindromic Repeats ID: 766114
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CRISPR and Viral Diagnostics Susan EibenJun HuangKira Slepchenko https://www.youtube.com/watch?v=22F85FOAyik
CRISPR-Cas9 https://labiotech.eu/features/crispr-cas9-review-gene-editing-tool/ Clusters of Regularly Interspaced Short Palindromic Repeats CRISPR associated protein 9 Natural bacterial viral defense
CRISPR - Cas based diagnostic Guide RNACRISPRCas12 or 13 Reporter Batista, Angelo C., and Luis GC Pacheco. "Detecting pathogens with Zinc-Finger, TALE and CRISPR-based programmable nucleic acid binding proteins." Journal of microbiological methods (2018).
https://blog.addgene.org/finding-nucleic-acids-with-sherlock-and-detectr Specific high-sensitivity enzymemaic reporter unlocking DNA Endonuclease Targeted CRISPR Trans Reporter
Lobato, I.M. et al, 2017. Recombinase polymerase amplification: Basics, applications and recent advances. TrAC Trends in Analytical Chemistry. cellular DNA synthesis recombination and repair RPA: Recombinase Polymerase Amplification
RPA: Recombinase Polymerase Amplification Rohrman, B.A. and Richards-Kortum, R.R., 2012. A paper and plastic device for performing recombinase polymerase amplification of HIV DNA. Lab on a chip , 12 (17), pp.3082-3088.
Cas12 and Cas13 Guide RNA/target recognitionspecific cut activates the enzyme collateral cutting https://phys.org/news/2018-02-crispr-based-technology-viral-dna.html
SHERLOCK (specific high-sensitivity enzymatic reporter unlocking)Myhrvold, C., Freije, C.A., Gootenberg, J.S., Abudayyeh, O.O., Metsky, H.C., Durbin, A.F., Kellner, M.J., Tan, A.L., Paul, L.M., Parham, L.A. and Garcia, K.F., 2018. Field-deployable viral diagnostics using CRISPR-Cas13. Science , 360(6387), pp.444-448.
Detecting Different Viruses Myhrvold, C., Freije, C.A., Gootenberg, J.S., Abudayyeh, O.O., Metsky, H.C., Durbin, A.F., Kellner, M.J., Tan, A.L., Paul, L.M., Parham, L.A. and Garcia, K.F., 2018. Field-deployable viral diagnostics using CRISPR-Cas13. Science, 360 (6387), pp.444-448.
Viral Strain Detection Pardee, K., Green, A.A., Takahashi, M.K., Braff, D., Lambert, G., Lee, J.W., Ferrante, T., Ma, D., Donghia, N., Fan, M. and Daringer, N.M., 2016. Rapid, low-cost detection of Zika virus using programmable biomolecular components. Cell , 165(5), pp.1255-1266.
Viral Strain Detection Gootenberg, J.S., Abudayyeh, O.O., Lee, J.W., Essletzbichler, P., Dy, A.J., Joung, J., Verdine, V., Donghia, N., Daringer, N.M., Freije, C.A. and Myhrvold, C., 2017. Nucleic acid detection with CRISPR-Cas13a/C2c2. Science , p.eaam9321.
The Importance/Impact In the field testing for virusesIsothermicLow cost Testing strips Dengue vs. Zika Tracking viral strains Location Virulence: symptoms, severity Inform epidemiological models Current methods are expensive, long development https://www.youtube.com/watch?v=22F85FOAyik
Summary CRISPR Cas based diagnostic similar to CRISPR Cas9 SHERLOCK and DETECTR Employs recombinant polymerase amplification (RPA) Cas12a and Cas13 Detection of different viruses and strains https://www.youtube.com/watch?v=22F85FOAyik
https://phys.org/news/2018-02-crispr-based-technology-viral-dna.html
Sources Batista, Angelo C., and Luis GC Pacheco. "Detecting pathogens with Zinc-Finger, TALE and CRISPR-based programmable nucleic acid binding proteins." Journal of microbiological methods (2018). Gootenberg, J.S., Abudayyeh, O.O., Lee, J.W., Essletzbichler, P., Dy, A.J., Joung, J., Verdine, V., Donghia, N., Daringer, N.M., Freije, C.A. and Myhrvold, C., 2017. Nucleic acid detection with CRISPR-Cas13a/C2c2. Science , p.eaam9321. Lobato, I.M. and O’Sullivan, C.K., 2017. Recombinase polymerase amplification: Basics, applications and recent advances. TrAC Trends in Analytical Chemistry . Myhrvold, C., Freije, C.A., Gootenberg, J.S., Abudayyeh, O.O., Metsky, H.C., Durbin, A.F., Kellner, M.J., Tan, A.L., Paul, L.M., Parham, L.A. and Garcia, K.F., 2018. Field-deployable viral diagnostics using CRISPR-Cas13. Science , 360 (6387), pp.444-448. Pardee, K., Green, A.A., Takahashi, M.K., Braff, D., Lambert, G., Lee, J.W., Ferrante, T., Ma, D., Donghia, N., Fan, M. and Daringer, N.M., 2016. Rapid, low-cost detection of Zika virus using programmable biomolecular components. Cell , 165 (5), pp.1255-1266. Rohrman, B.A. and Richards-Kortum, R.R., 2012. A paper and plastic device for performing recombinase polymerase amplification of HIV DNA. Lab on a chip , 12 (17), pp.3082-3088. https://blog.addgene.org/finding-nucleic-acids-with-sherlock-and-detectr https://labiotech.eu/features/crispr-cas9-review-gene-editing-tool/ https://phys.org/news/2018-02-crispr-based-technology-viral-dna.html https://www.youtube.com/watch?v=22F85FOAyik