Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potential
The scientific and technological advent of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is one of the most exciting developments of the past decade, particularly in the field of gene editing. The technology has two essential components, (1) a guide RNA to match a targeted gene...
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| Format: | Article |
| Language: | English |
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Cambridge University Press
2022-01-01
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| Series: | QRB Discovery |
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| Online Access: | https://www.cambridge.org/core/product/identifier/S2633289222000072/type/journal_article |
| _version_ | 1811155725641056256 |
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| author | Juan G. Santiago |
| author_facet | Juan G. Santiago |
| author_sort | Juan G. Santiago |
| collection | DOAJ |
| description | The scientific and technological advent of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is one of the most exciting developments of the past decade, particularly in the field of gene editing. The technology has two essential components, (1) a guide RNA to match a targeted gene and (2) a CRISPR-associated protein (e.g. Cas 9, Cas12 or Cas13) that acts as an endonuclease to specifically cut DNA. This specificity and reconfigurable nature of CRISPR has also spurred intense academic and commercial interest in the development of CRISPR-based molecular diagnostics. CRISPR Cas12 and Cas13 orthologs are most commonly applied to diagnostics, and these cleave and become activated by DNA and RNA targets, respectively. Despite the intense research interest, the limits of detection (LoDs) and applications of CRISP-based diagnostics remain an open question. A major reason for this is that reports of kinetic rates have been widely inconsistent, and the vast majority of these reports contain gross errors including violations of basic conservation and kinetic rate laws. It is the intent of this Perspective to bring attention to these issues and to identify potential improvements in the manner in which CRISPR kinetic rates and assay LoDs are reported and compared. The CRISPR field would benefit from verifications of self-consistency of data, providing sufficient data for reproduction of experiments, and, in the case of reports of novel assay LoDs, concurrent reporting of the associated kinetic rate constants. The early development of CRISPR-based diagnostics calls for self-reflection and urges us to proceed with caution. |
| first_indexed | 2024-04-10T04:38:24Z |
| format | Article |
| id | doaj.art-329f5bbcb9dc45b98bbe73b0b8873ee3 |
| institution | Directory Open Access Journal |
| issn | 2633-2892 |
| language | English |
| last_indexed | 2024-04-10T04:38:24Z |
| publishDate | 2022-01-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | QRB Discovery |
| spelling | doaj.art-329f5bbcb9dc45b98bbe73b0b8873ee32023-03-09T12:43:36ZengCambridge University PressQRB Discovery2633-28922022-01-01310.1017/qrd.2022.7Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potentialJuan G. Santiago0https://orcid.org/0000-0001-8652-5411Department of Mechanical Engineering, Stanford University, Stanford, CA 94305, USAThe scientific and technological advent of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is one of the most exciting developments of the past decade, particularly in the field of gene editing. The technology has two essential components, (1) a guide RNA to match a targeted gene and (2) a CRISPR-associated protein (e.g. Cas 9, Cas12 or Cas13) that acts as an endonuclease to specifically cut DNA. This specificity and reconfigurable nature of CRISPR has also spurred intense academic and commercial interest in the development of CRISPR-based molecular diagnostics. CRISPR Cas12 and Cas13 orthologs are most commonly applied to diagnostics, and these cleave and become activated by DNA and RNA targets, respectively. Despite the intense research interest, the limits of detection (LoDs) and applications of CRISP-based diagnostics remain an open question. A major reason for this is that reports of kinetic rates have been widely inconsistent, and the vast majority of these reports contain gross errors including violations of basic conservation and kinetic rate laws. It is the intent of this Perspective to bring attention to these issues and to identify potential improvements in the manner in which CRISPR kinetic rates and assay LoDs are reported and compared. The CRISPR field would benefit from verifications of self-consistency of data, providing sufficient data for reproduction of experiments, and, in the case of reports of novel assay LoDs, concurrent reporting of the associated kinetic rate constants. The early development of CRISPR-based diagnostics calls for self-reflection and urges us to proceed with caution.https://www.cambridge.org/core/product/identifier/S2633289222000072/type/journal_articleClustered Regularly Interspaced Short Palindromic RepeatsCRISPRCRISPR-Casenzyme kineticslimit of detectionmolecular diagnostics |
| spellingShingle | Juan G. Santiago Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potential QRB Discovery Clustered Regularly Interspaced Short Palindromic Repeats CRISPR CRISPR-Cas enzyme kinetics limit of detection molecular diagnostics |
| title | Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potential |
| title_full | Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potential |
| title_fullStr | Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potential |
| title_full_unstemmed | Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potential |
| title_short | Inconsistent treatments of the kinetics of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) impair assessment of its diagnostic potential |
| title_sort | inconsistent treatments of the kinetics of clustered regularly interspaced short palindromic repeats crispr impair assessment of its diagnostic potential |
| topic | Clustered Regularly Interspaced Short Palindromic Repeats CRISPR CRISPR-Cas enzyme kinetics limit of detection molecular diagnostics |
| url | https://www.cambridge.org/core/product/identifier/S2633289222000072/type/journal_article |
| work_keys_str_mv | AT juangsantiago inconsistenttreatmentsofthekineticsofclusteredregularlyinterspacedshortpalindromicrepeatscrisprimpairassessmentofitsdiagnosticpotential |