PT  - JOURNAL ARTICLE
AU  - Linxian Li
AU  - Shiyuan Li
AU  - Jin Wang
TI  - CRISPR-Cas12b-assisted nucleic acid detection platform
AID  - 10.1101/362889
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 362889
4099  - http://biorxiv.org/content/early/2018/07/06/362889.short
4100  - http://biorxiv.org/content/early/2018/07/06/362889.full
AB  - Rapid molecular diagnostic technology is very useful in many areas, including public health, environmental testing and criminal investigation. We recently showed that Cas12a had trans-cleavage activity upon collateral single-stranded DNA (ssDNA), with which the HOLMES platform (one-HOur Low-cost Multipurpose highly Efficient System) was developed. Here, we combine the thermophilic Cas12b, which also has the ssDNA trans-cleavage activity, with Loop-Mediated Isothermal Amplification (LAMP), and create HOLMESv2. In HOLMESv2, LAMP amplification and Cas12b trans-cleavage can be integrated into a one-step system with a constant temperature, which therefore brings much convenience in nucleic acid detection. Moreover, we also simplify the RNA detection procedures in HOLMESv2, using an RNA-dependent DNA polymerase for amplification and therefore omitting an extra reverse transcription step.One Sentence Summary We combine LAMP and Cas12b to develop HOLMESv2 for conveniently detecting target nucleic acid in a one-step approach.