| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | cas12a |
| Uniprot No | U2UMQ6 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-1307aa |
| 氨基酸序列 | MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLNLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLRNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGKITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLKKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKARNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPLEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFHVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRSLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVVVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDVKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENHRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMRNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESKDLKLQNGISNQDWLAYIQELRN |
| 预测分子量 | 158.6 kDa |
| 蛋白标签 | His tag N-Terminus |
| 缓冲液 | PBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300. |
| 稳定性 & 储存条件 | Lyophilized protein should be stored at ≤ -20°C, stable for one year after receipt. Reconstituted protein solution can be stored at 2-8°C for 2-7 days. Aliquots of reconstituted samples are stable at ≤ -20°C for 3 months. |
| 复溶 | Always centrifuge tubes before opening.Do not mix by vortex or pipetting. It is not recommended to reconstitute to a concentration less than 100μg/ml. Dissolve the lyophilized protein in distilled water. Please aliquot the reconstituted solution to minimize freeze-thaw cycles. |
以下是关于Cas12a重组蛋白的3-4篇代表性文献，包含文献名称、作者及摘要概括：
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1. **文献名称**: *Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System*
**作者**: Zetsche, B., et al. (包括张锋、Doudna等)
**摘要**: 该研究首次鉴定Cas12a(Cpf1)为CRISPR-Cas系统的新型核酸酶，证明其通过单一RNA引导切割双链DNA，并揭示其与Cas9不同的特性(如交错切割、无需tracrRNA)，为基因编辑工具开发奠定基础。
2. **文献名称**: *CRISPR-Cas12a target binding unleashes single-stranded DNase activity*
**作者**: Chen, J.S., et al.
**摘要**: 本文解析Cas12a在结合靶标DNA后激活非特异性单链DNA反式切割活性的机制，并基于此开发了DETECTR诊断技术，利用重组Cas12a蛋白实现高灵敏度核酸检测，应用于HPV分型等临床检测。
3. **文献名称**: *Engineered CRISPR-Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing*
**作者**: Li, S.Y., et al.
**摘要**: 通过工程化改造重组Cas12a蛋白，增强其在哺乳动物细胞中的基因编辑效率，并拓展其靶向范围，同时开发了基于Cas12a的碱基编辑系统，为治疗性基因编辑提供优化工具。
4. **文献名称**: *DNA interrogation by the CRISPR RNA-guided endonuclease Cas12a*
**作者**: Swarts, D.C., et al.
**摘要**: 研究利用重组Cas12a蛋白探究其DNA识别机制，发现其对富含胸腺嘧啶的PAM序列偏好性，并揭示其反式切割活性在体外诊断中的应用潜力，为后续分子诊断技术开发提供理论支持。
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以上文献涵盖了Cas12a重组蛋白的机制解析、基因编辑优化及诊断应用等方向，均为该领域的重要研究。
Cas12a (formerly Cpf1) is a RNA-guided DNA endonuclease belonging to the Class 2 Type V CRISPR-Cas system, first characterized in 2015. Unlike the widely used Cas9. Cas12a recognizes a thymidine-rich protospacer adjacent motif (PAM: 5'-TTTV-3') and generates staggered DNA double-strand breaks with cohesive overhangs, offering advantages in precise gene insertion. It processes its own CRISPR RNA (crRNA) from a precursor array without requiring a trans-activating crRNA (tracrRNA), simplifying multiplex genome editing by enabling expression of multiple guide RNAs from a single transcript.
Recombinant Cas12a proteins are engineered for experimental and therapeutic applications through heterologous expression in bacterial systems (e.g., *E. coli*). Purification typically involves affinity chromatography tags (His-tag, GST-tag) followed by buffer optimization to maintain enzymatic activity. Structural studies reveal a bilobed architecture with a nuclease lobe (Nuc) and α-helical recognition lobe (Rec), enabling target DNA unwinding and cleavage via a single catalytic site (RuvC domain).
Cas12a exhibits off-target activity lower than Cas9 in some contexts and demonstrates collateral cleavage of single-stranded DNA upon target recognition (trans-nuclease activity), which has been exploited in diagnostic tools like SHERLOCK and DETECTR. Engineered variants (e.g., enAsCas12a, HypaCas12a) with enhanced specificity or expanded PAM compatibility (e.g., NG, GAA) have been developed through directed evolution. Its smaller size compared to SpCas9 facilitates delivery via adeno-associated virus (AAV) vectors for *in vivo* applications. Cas12a-based systems are increasingly used in plant genome editing, antimicrobial agent development, and molecular diagnostics, highlighting its versatility beyond traditional CRISPR-Cas9 platforms.
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