![]() Lowder LG, Zhou J, Zhang Y et al (2018) Robust transcriptional activation in plants using multiplexed CRISPR-Act2.0 and mTALE-Act systems. Pan C, Sretenovic S, Qi Y (2021) CRISPR/dCas-mediated transcriptional and epigenetic regulation in plants. Key wordsĭominguez AA, Lim WA, Qi LS (2016) Beyond editing: repurposing CRISPR-Cas9 for precision genome regulation and interrogation. We present detailed procedures on the sgRNA design, CRISPR–Act3.0 system construction, Agrobacterium-mediated transformation of Arabidopsis using the floral dip method, and identification of desired transgenic plants. In this chapter, we provide a step-by-step guide for efficient gene activation in Arabidopsis based on a highly robust CRISPRa system, CRISPR–Act3.0. The CRISPRa systems enable precise, scalable, and robust RNA-guided transcription activation, holding great potential for a variety of fundamental and translational research. CRISPR activation (CRISPRa) represents a novel gain-of-function (GOF) strategy, conferring robust over-expression of the target gene within its native chromosomal context. Beyond genome editing, CRISPR/Cas systems, based on nuclease-deficient Cas9 (dCas9), have been repurposed as an RNA-guided platform for transcriptional regulation. ![]() PaeR7I does not recognize the sequence CTCTCGAG.Įfficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.The CRISPR/Cas system has emerged as a versatile platform for sequence-specific genome engineering in plants. ![]() Prolonged incubation with NdeI may lead to removal of additional nucleotides. ![]() Sticky ends from different SexAI sites may not be compatible. Sticky ends from different CsiI sites may not be compatible. Sticky ends from different SfiI sites may not be compatible. ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |