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1.
Plast Reconstr Surg ; 2024 Jul 22.
Article in English | MEDLINE | ID: mdl-39046832

ABSTRACT

PURPOSE: To evaluate whether the sequence of osteotomies influences the accuracy of maxillary positioning in patients with cleft palate ± cleft lip undergoing bimaxillary orthognathic surgery (OGS). METHODS: This was a prospective study of patients with Veau 2-4 clefts who underwent bimaxillary OGS at tertiary-care children's hospital over a 3-year period. The primary predictor variable was the sequence of osteotomies (maxilla-first versus mandible-first). The primary outcome of interest was the concordance between the planned and achieved maxillary position, as assessed using linear and angular measurements. Secondary study predictors were demographic and surgical variables. Differences between groups were compared using non-parametric independent samples tests for continuous measures (data reported as median and interquartile range, IQR) and chi-squared tests for categorical measures. For all analyses, p≤ 0.05 was considered significant. RESULTS: Subjects who underwent maxilla-first (n=15) and mandible-first (n=16) operations were comparable with regard to age, gender, cleft type, skeletal classification, segmental maxillary osteotomy, and magnitude of maxillary movement (p ≥ 0.09). The planned sagittal and vertical positions of the maxilla were similarly accurate between the two groups (p ≥ 0.68). Angular accuracy was also comparable (p ≥ 0.56) between the study groups. CONCLUSION: In patients with CP ± CL undergoing bimaxillary orthognathic surgery, use of mandible-first sequencing, when compared to maxilla-first sequencing, does not impact accuracy of maxillary positioning in the immediate post-operative period in well-selected patients.

3.
Cleft Palate Craniofac J ; 60(5): 639-644, 2023 05.
Article in English | MEDLINE | ID: mdl-35044260

ABSTRACT

This study sought to identify disparities in the timing of alveolar bone grafting (ABG) surgery and the replacement strategy for missing maxillary lateral incisors for patients with clefts.A retrospective record review identified patients who underwent ABG. Multivariable regression analyzed the independent contribution of each variable.This institutional study was performed at the University of California, San Francisco.Patients who presented under age 12 and underwent secondary ABG between 2012 and 2020 (n = 160).The age at secondary ABG and the recommended dental replacement treatment for each patient, either dental implantation or canine substitution.The average age at ABG was 10.8 ± 2.1 years, 106 (66.3%) patients were not White, and 80 (50.0%) had private insurance. Independent predictors of older age at ABG included an income below $ 50 000 as estimated from ZIP code (ß = 15.0 months, 95% CI, 5.7-24.3, P = .002) and identifying as a race other than White (ß = 10.1 months, 95% CI, 2.1-18.0, P = .01). After ABG, patients were more likely to undergo dental implantation over canine substitution if they were female (odds ratio [OR] = 4.3, 95% CI, 1.3-17.1, P = .02) or had private insurance (OR = 12.5, 95% CI, 2.2-143.2, P = .01).Patients who were low-income or not White experienced delays in ABG, whereas dental implantation was more likely to be recommended for patients with private insurance. Understanding the sources of disparities in dental reconstruction of cleft deformities may reveal opportunities to improve equity.


Subject(s)
Alveolar Bone Grafting , Cleft Lip , Cleft Palate , Female , Male , Humans , Cleft Palate/surgery , Cleft Lip/surgery , Retrospective Studies , Incisor , Bone Transplantation
4.
J Craniofac Surg ; 34(3): 1010-1014, 2023 May 01.
Article in English | MEDLINE | ID: mdl-36210502

ABSTRACT

BACKGROUND: Facial feminization surgery (FFS) remains inaccessible to many transgender patients. Zuckerberg San Francisco General Hospital (ZSFG) was among the first public, safety-net hospitals to perform FFS. The purpose of this study is to examine the postoperative outcomes of patients who underwent FFS at ZSFG and describe barriers to providing FFS in a public hospital setting. METHODS: A retrospective review identified patients who underwent FFS at ZSFG. Demographic data, comorbidity profiles, postoperative outcomes, and hospital utilization data were collected from the medical records. FACE-Q modules (scored 0-100) were used to survey patient satisfaction at least 1 year postoperatively. RESULTS: Seventeen patients underwent comprehensive FFS surgery at ZSFG. The median age was 41 years [interquartile range (IQR): 38-55], median body mass index was 26.4 (IQR: 24.1-31.3). Patients underwent a median of 9 procedures, the most common of which included frontal cranioplasty (n=13, 77%), open brow lift (n=13, 77%), rhinoplasty (n=12, 71%), and mandible contouring (n=12, 71%). There were no complications, readmissions, or reoperations within 30 days. Patients reported high satisfaction with the surgical outcome (median: 87, IQR: 87-100), excellent postoperative psychological functioning (median: 100, IQR: 88-100), and low levels of appearance-related distress (median: 3, IQR: 0-35). An estimated 243 operating room hours and 51 inpatient bed days were required to cover all FFS procedures. CONCLUSIONS: Performing FFS in a public, safety-net hospital was associated with zero postoperative complications, few revision procedures, and excellent patient satisfaction. Limited operating room hours and inpatient availability represented barriers to providing FFS in this setting.


Subject(s)
Face , Sex Reassignment Surgery , Male , Humans , Adult , Face/surgery , Safety-net Providers , Feminization/surgery , Esthetics, Dental
5.
J Craniofac Surg ; 33(8): 2422-2426, 2022.
Article in English | MEDLINE | ID: mdl-36409867

ABSTRACT

The purpose of this study was to identify racial and socioeconomic disparities in craniosynostosis evaluation and treatment, from referral to surgery. Patients diagnosed with craniosynostosis between 2012 and 2020 at a single center were identified. Chart review was used to collect demographic variables, age at referral to craniofacial care, age at diagnosis, age at surgery, and surgical technique (open versus limited incision). Multivariable linear and logistic regression models with lasso regularization assessed the independent effect of each variable. A total of 298 patients were included. Medicaid insurance was independently associated with a delay in referral of 83 days [95% confidence interval (CI) 4-161, P=0.04]. After referral, patients were diagnosed a median of 21 days later (interquartile range 7-40), though this was significantly prolonged in patients who were not White (ß 23 d, 95% CI 9-38, P=0.002), had coronal synostosis (ß 24 d, 95% CI 2-46, P=0.03), and had multiple suture synostosis (ß 47 d, 95% CI 27-67, P<0.001). Medicaid insurance was also independently associated with diagnosis over 3 months of age (risk ratio 1.3, 95% CI 1.1-1.4, P=0.002) and undergoing surgery over 1 year of age (risk ratio 3.9, 95% CI 1.1-9.4, P=0.04). In conclusion, Medicaid insurance was associated with a 3-month delay in referral to craniofacial specialists and increased risk of diagnosis over 3 months of age, limiting surgical treatment options in this group. Patients with Medicaid also faced a 4-fold greater risk of delayed surgery, which could result in neurodevelopmental sequelae.


Subject(s)
Craniosynostoses , Healthcare Disparities , United States , Humans , Racial Groups , Craniosynostoses/diagnosis , Craniosynostoses/surgery , Medicaid , Socioeconomic Factors
6.
J Craniofac Surg ; 33(4): e443-e445, 2022 Jun 01.
Article in English | MEDLINE | ID: mdl-36041099

ABSTRACT

ABSTRACT: Pediatric cranioplasty is indicated to repair skull defects with a wide variety of etiologies. The choice of graft material used to fill the defect is of paramount importance to the long-term success of this procedure. A variety of synthetic products have been commercially developed to avoid donor site morbidity. Here, the authors present the case of a 13-year-old boy with cranial Langerhans cell histiocytosis who underwent cranioplasty with a novel, calcium phosphate-based bone graft substitute (Montage). The patient presented 2 years postoperatively with a foreign body giant cell reaction that required explantation of the graft. The authors discuss potential considerations in choosing the most appropriate graft, potential contributors to this late adverse outcome, and the need for further research into the use of novel allograft materials in pediatric cranioplasty.


Subject(s)
Bone Substitutes , Plastic Surgery Procedures , Adolescent , Bone Transplantation/methods , Foreign-Body Reaction/etiology , Foreign-Body Reaction/surgery , Humans , Male , Plastic Surgery Procedures/methods , Retrospective Studies , Skull/surgery
8.
Plast Reconstr Surg Glob Open ; 10(2): e4097, 2022 Feb.
Article in English | MEDLINE | ID: mdl-35169528

ABSTRACT

BACKGROUND: Strip craniectomy with orthotic helmet therapy (SCOT) is an increasingly supported treatment for metopic craniosynostosis, although the long-term efficacy of deformity correction remains poorly defined. We compared the longterm outcomes of SCOT versus open cranial vault reconstruction (OCVR). METHODS: Patients who underwent OCVR or SCOT for isolated metopic synostosis with at least 3 years of follow-up were identified at our institution. Anthropometric measurements were used to assess baseline severity and postoperative skull morphology. Independent laypersons and craniofacial surgeons rated the appearance of each patient's 3D photographs, compared to normal controls. RESULTS: Thirty-five patients were included (15 SCOT and 20 OCVR), with similar follow-up between groups (SCOT 7.9 ± 3.2 years, OCVR 9.2 ± 4.1 years). Baseline severity and postoperative anthropometric measurements were equivalent. Independent adolescent raters reported that the forehead, eye, and overall appearance of SCOT patients was better than OCVR patients (P < 0.05, all comparisons). Craniofacial surgeons assigned Whitaker class I to a greater proportion of SCOT patients with moderate-to-severe synostosis (72.2 ± 5.6%) compared with OCVR patients with the same severity (33.3 ± 9.2%, P = 0.02). Parents of children who underwent SCOT reported equivalent satisfaction with the results of surgery (100% versus 95%, P > 0.99), and were no more likely to report bullying (7% versus 15%, P = 0.82). CONCLUSIONS: SCOT was associated with superior long-term appearance and perioperative outcomes compared with OCVR. These findings suggest that SCOT should be the treatment of choice for patients with a timely diagnosis of metopic craniosynostosis.

9.
J Surg Educ ; 79(1): 20-24, 2022.
Article in English | MEDLINE | ID: mdl-34446382

ABSTRACT

OBJECTIVE: The COVID-19 pandemic provided an opportunity for surgical residency programs to rethink their methods of evaluating and recruiting candidates. However, the past year has not been seamless, with a soaring number of applications, reports of programs and applicants having difficulty evaluating each other, and an increasingly uneven distribution of interviews among applicants. Consequently, many have called for national changes to the residency application process to address these longstanding concerns. RESULTS: Here, we review the evolving literature and advocate for the permanent adoption of visiting rotations, virtual interviews with a universal release date and data-driven attendance limits, and opportunities for in-person applicant visits. CONCLUSIONS: We believe these changes leverage the strengths of each format, allow for satisfactory bidirectional evaluation, and promote principles of justice, equity, diversity, and inclusion.


Subject(s)
COVID-19 , Internship and Residency , Humans , Pandemics , SARS-CoV-2 , Students
10.
Plast Reconstr Surg Glob Open ; 9(1): e3351, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33564582

ABSTRACT

Whether treatment of cleft palate (CP) associated with Robin sequence (RS) should attain outcomes similar to those of isolated cleft palate (ICP) remains unknown. This study compares treatment and outcomes in both conditions and delineates predictors of long-term outcome. METHODS: This retrospective case series of consecutive syndromic and isolated RS- and ICP-patients (1990-2016) includes indications and outcomes of straight-line repair with intravelar veloplasty (SLIV) or Furlow repair depending on cleft and airway characteristics. RESULTS: Seventy-five RS and 83 ICP patients underwent CP repair. Velopharyngeal insufficiency (VPI) occurred in 41% of RS versus 17% of ICP patients (P = 0.012), and in 60% of patients with syndromic RS versus 16% with isolated RS (P = 0.005). In multivariable logistic regression analysis, wider and more severe CP anatomy was the only factor independently associated with VPI (P = 0.028), in contrast to age at repair, syndromic RS compared with isolated RS, and isolated RS compared with ICP and initial tongue-lip adhesion. Secondary Furlow after primary SLIV was used to treat VPI in all groups, and more frequently in syndromic versus isolated RS patients (P = 0.025). CONCLUSIONS: Variability of RS anatomy and airway compromise necessitates individualized treatment protocols. Despite differing CP etiology and other variables, our findings demonstrate cleft anatomy as the only independent variable predictive of VPI comparing RS and ICP patients. Patients with isolated RS should ultimately attain similar VPI outcomes compared with ICP patients. Obstructive speech operations in RS patients can be avoided without compromising speech outcome by reserving the prsocedure for secondary cases.

11.
Neurosurgery ; 88(6): 1088-1094, 2021 05 13.
Article in English | MEDLINE | ID: mdl-33575788

ABSTRACT

BACKGROUND: Posterior cervical decompression and fusion (PCF) is a common procedure used to treat various cervical spine pathologies, but the 90-d outcomes following PCF surgery continue to be incompletely defined. OBJECTIVE: To identify risk factors associated with 90-d readmission and reoperation following PCF surgery. METHODS: Adults undergoing PCF from 2012 to 2020 were identified. Demographic and radiographic data, surgical characteristics, and 90-d outcomes were collected. Univariate analysis was performed using Student's t-test, chi square, and Fisher exact tests as appropriate. Multivariable logistic regression models with lasso penalty were used to analyze various risk factors. RESULTS: A total of 259 patients were included. The 90-d readmission and reoperation rates were 9.3% and 4.6%, respectively. The most common reason for readmission was surgical site infection (SSI) (33.3%) followed by new neurological deficits (16.7%). Patients who smoked tobacco had 3-fold greater odds of readmission compared to nonsmokers (odds ratio [OR]: 3.48; 95% CI 1.87-6.67; P = .0001). Likewise, the most common reason for reoperation was SSI (33.3%) followed by seroma and implant failure (25.0% each). Smoking was also an independent risk factor for reoperation, associated with nearly 4-fold greater odds of return to the operating room (OR: 3.53; 95% CI 1.53-8.57; P = .003). CONCLUSION: Smoking is a significant predictor of 90-d readmission and reoperation in patients undergoing PCF surgery. Smoking cessation should be strongly considered preoperatively in elective PCF cases to minimize the risk of 90-d readmission and reoperation.


Subject(s)
Cervical Vertebrae/surgery , Decompression, Surgical/statistics & numerical data , Patient Readmission/statistics & numerical data , Reoperation/statistics & numerical data , Smoking/epidemiology , Spinal Fusion/statistics & numerical data , Adult , Aged , Humans , Male , Middle Aged , Retrospective Studies , Risk Factors , Second-Look Surgery , Surgical Wound Infection/etiology
12.
AJR Am J Roentgenol ; 217(3): 709-717, 2021 09.
Article in English | MEDLINE | ID: mdl-33377802

ABSTRACT

Facial feminization surgery is an increasingly performed component of gender affirmation surgery for transgender women. Preoperative facial CT is performed to plan the adjustment of the patient's masculine characteristics to feminine and to plan operative navigation around specific readily identifiable anatomic structures. In the upper face, surgery is performed to reduce the prominence of the brow and increase the nasofrontal angle; the radiology report should indicate the frontal sinus and supraorbital foramen anatomy. In the midface, rhinoplasty is performed to increase the nasofrontal and nasolabial angles; the radiology report should indicate the presence of a dorsal hump and septal deviation or spurring. In the lower face, the prominence of the chin and squareness of the jaw are adjusted via genioplasty and mandible contouring, respectively; the radiology report should describe the location and potential anatomic variations of the inferior alveolar nerve and mental foramina as well as the presence of dental abnormalities that directly inform the surgical approach. CT may also be performed if there is clinical suspicion for postoperative complications such as hardware fracture or osteotomy through the supraorbital or mental foramen. Familiarity with these findings will facilitate improved communication between radiologists and surgeons, thereby contributing to the care of transgender women.


Subject(s)
Facial Bones/diagnostic imaging , Facial Bones/surgery , Feminization/surgery , Postoperative Complications/diagnostic imaging , Preoperative Care/methods , Tomography, X-Ray Computed/methods , Adult , Face/surgery , Female , Humans , Male , Transsexualism/surgery
13.
AJR Am J Roentgenol ; 2020 Dec 30.
Article in English | MEDLINE | ID: mdl-33377414

ABSTRACT

Facial feminization surgery (FFS) is an increasingly performed component of gender affirmation surgery for transgender women. Preoperative facial CT is performed to plan the adjustment of the patient's masculine characteristics to feminine, and to plan operative navigation around specific readily identifiable anatomic structures. In the upper face, surgery is performed to reduce the prominence of the brow and increase the nasofrontal angle; the radiology report should indicate the frontal sinus and supraorbital foramen anatomy. In the midface, rhinoplasty is performed to increase the nasofrontal and nasolabial angles; the radiology report should indicate presence of a dorsal hump and septal deviation or spurring. In the lower face, prominence of the chin and squareness of the jaw are adjusted via genioplasty and mandible contouring, respectively; the radiology report should describe the location and potential anatomic variations of the inferior alveolar nerve and mental foramina, as well as presence of dental abnormalities that directly inform the surgical approach. CT may also be performed if there is clinical suspicion for postoperative complications such as hardware fraction or osteotomy through the supraorbital or mental foramen. Familiarity with these findings will facilitate improved communication between radiologists and surgeons, thereby contributing to the care of transgender women.

14.
Plast Reconstr Surg Glob Open ; 8(9): e3143, 2020 Sep.
Article in English | MEDLINE | ID: mdl-33133981

ABSTRACT

Craniofacial clinics are composed of multidisciplinary teams of providers to deliver coordinated and comprehensive patient care. The coronavirus disease of 2019 (COVID-19) pandemic has disrupted this model, as social distancing guidelines have precluded in-person patient appointments and forced clinics to reconsider their method of care delivery. The University of California, San Francisco, Craniofacial Center has continued to serve patients during this acute period, adopting a hybrid model in which the vast majority of patients are seen through telehealth and a limited number of patients are evaluated in-person. Surveyed patients and families reported high rates of satisfaction, with time savings cited as a particular benefit. Furthermore, most felt comfortable using the video technology required for their appointment. This experience has demonstrated to us that multidisciplinary craniofacial evaluations can be effectively delivered in a telehealth format and has informed our conception of idealized clinic structure. Moving forward, we intend to utilize telehealth visits for selected components of craniofacial evaluations in an effort to maximize efficiency and minimize burden, including addressing barriers to accessing care. Benefits of a hybrid model will include decongestion of clinics and waiting areas, allowing social distancing, addressing clinic space limits, and increased efficiency by eliminating the need for patient and family movement. Demonstration of the safety and efficacy of telehealth visits, combined with regulatory reform that improves reimbursement and allows for appointments across state lines, will be critical for this model to persist beyond the pandemic.

15.
Neurosurgery ; 87(5): 1016-1024, 2020 10 15.
Article in English | MEDLINE | ID: mdl-32577734

ABSTRACT

BACKGROUND: For laminectomy and posterior spinal fusion (LPSF) surgery for cervical spondylotic myelopathy (CSM), the evidence is unclear as to whether fusions should cross the cervicothoracic junction (CTJ). OBJECTIVE: To compare LPSF outcomes between those with and without lower instrumented vertebrae (LIV) crossing the CTJ. METHODS: A consecutive series of adults undergoing LPSF for CSM from 2012 to 2018 with a minimum of 12-mo follow-up were identified. LPSF with subaxial upper instrumented vertebrae and LIV between C6 and T2 were included. Clinical and radiographic outcomes were compared. RESULTS: A total of 79 patients were included: 46 crossed the CTJ (crossed-CTJ) and 33 did not. The mean follow-up was 22.2 mo (minimum: 12 mo). Crossed-CTJ had higher preoperative C2-7 sagittal vertical axis (cSVA) (33.3 ± 16.0 vs 23.8 ± 12.4 mm, P = .01) but similar preoperative cervical lordosis (CL) and CL minus T1-slope (CL minus T1-slope) (P > .05, both comparisons). The overall reoperation rate was 3.8% (crossed-CTJ: 2.2% vs not-crossed: 6.1%, P = .37). In adjusted analyses, crossed-CTJ was associated with superior cSVA (ß = -9.7; P = .002), CL (ß = 6.2; P = .04), and CL minus T1-slope (ß = -6.6; P = .04), but longer operative times (ß = 46.3; P = .001). Crossed- and not-crossed CTJ achieved similar postoperative patient-reported outcomes [Visual Analog Scale (VAS) neck pain, VAS arm pain, Nurick Grade, Modified Japanese Orthopedic Association Scale, Neck Disability Index, and EuroQol-5D] in adjusted multivariable analyses (adjusted P > .05). For the entire cohort, higher postoperative CL was associated with lower postoperative arm pain (adjusted Pearson's r -0.1, P = .02). No postoperative cervical radiographic parameters were associated with neck pain (P > .05). CONCLUSION: Subaxial LPSF for CSM that crossed the CTJ were associated with superior radiographic outcomes for cSVA, CL, and CL minus T1-slope, but longer operative times. There were no differences in neck pain or reoperation rate.


Subject(s)
Cervical Vertebrae/surgery , Laminectomy/methods , Spinal Cord Diseases/surgery , Spinal Fusion/methods , Thoracic Vertebrae/surgery , Adult , Aged , Cohort Studies , Female , Humans , Male , Middle Aged , Postoperative Complications/epidemiology , Postoperative Complications/etiology , Spinal Fusion/adverse effects , Treatment Outcome
16.
J Spine Surg ; 6(1): 323-333, 2020 Mar.
Article in English | MEDLINE | ID: mdl-32309669

ABSTRACT

Posterior cervical decompression and fusion (PCF) is a common surgical technique used to treat various cervical spine pathologies. However, there are various complications associated with PCF that can negatively impact patient outcome. We performed a comprehensive literature review to identify the most common complications following PCF using PubMed, Cochrane Database of Systematic Reviews, and Google Scholar. The overall complication rates of PCF are estimated to range from about 15% to 25% in the current literature. The most common immediate complications include acute blood loss anemia, surgical site infection (SSI), C5 palsy, and incidental durotomy; the most common long-term complications include adjacent segment degeneration, junctional kyphosis, and pseudoarthrosis. Three principal mechanisms are thought to contribute to complications. First, higher number of fusion levels, obesity, and more complex pathologies can increase the invasiveness of the planned procedure, thus increase complications. Second, wound healing and arthrodesis may be impaired due to poor blood flow due to various patient factors such as smoking, diabetes, increased frailty, steroid use, and other medical comorbidities. Finally, increased biomechanical stress on the upper instrumented vertebra (UIV) and lowest instrumented vertebra (LIV) may predispose patient to chronic degeneration and result in adjacent level degeneration and/or junctional problems. Reducing the modifiable risk factors pre-operatively can decrease the overall complication rate. Neurologic deficits may be reduced with adequate intraoperative decompression of neural elements. SSI may be reduced with meticulous wound closure that minimizes dead space, drain placement, and the use of intra-wound antibiotics. Careful design of the fusion construct with consideration in spinal alignment and biomechanics can help to reduce the rate of junctional problems. Spine surgeons should be aware of these complications associated with PCF and the corresponding prevention strategies optimize patient outcomes.

17.
Neurospine ; 16(3): 548-557, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31607087

ABSTRACT

OBJECTIVE: This retrospective cohort study seeks to identify risk factors associated with complications following posterior cervical laminectomy and fusion (PCLF) surgery. METHODS: Adults undergoing PCLF from 2012 through 2018 at a single center were identified. Demographic and radiographic data, surgical characteristics, and complication rates were compared. Multivariate logistic regression models identified independent predictors of complications following surgery. RESULTS: A total of 196 patients met the inclusion criteria and were included in the study. The medical, surgical, and overall complication rates were 10.2%, 23.0%, and 29.1% respectively. Risk factors associated with medical complications in multivariate analysis included impaired ambulation status (odds ratio [OR], 2.27; p=0.02) and estimated blood loss over 500 mL (OR, 3.67; p=0.02). Multivariate analysis revealed preoperative narcotic use (OR, 2.43; p=0.02) and operative time (OR, 1.005; p=0.03) as risk factors for surgical complication, whereas antidepressant use was a protective factor (OR, 0.21; p=0.01). Overall complication was associated with preoperative narcotic use (OR, 1.97; p=0.04) and higher intraoperative blood loss (OR, 1.0007; p=0.03). CONCLUSION: Preoperative narcotic use and estimated blood loss predicted the incidence of complications following PCLF for CSM. Ambulation status was a significant predictor of the development of a medical complication specifically. These results may help surgeons in counseling patients who may be at increased risk of complication following surgery.

18.
Nat Methods ; 15(7): 523-526, 2018 07.
Article in English | MEDLINE | ID: mdl-29967496

ABSTRACT

Robust approaches for chemogenetic control of protein function would have many biological applications. We developed stabilizable polypeptide linkages (StaPLs) based on hepatitis C virus protease. StaPLs undergo autoproteolysis to cleave proteins by default, whereas protease inhibitors prevent cleavage and preserve protein function. We created StaPLs responsive to different clinically approved drugs to bidirectionally control transcription with zinc-finger-based effectors, and used StaPLs to create single-chain, drug-stabilizable variants of CRISPR-Cas9 and caspase-9.


Subject(s)
Gene Expression Regulation/drug effects , Protein Engineering , CRISPR-Cas Systems , Dimerization , Gene Targeting , HEK293 Cells , HeLa Cells , Humans , Plasmids , Protein Folding , Viral Nonstructural Proteins/metabolism , Zinc Fingers
19.
Nat Methods ; 13(12): 993-996, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27798610

ABSTRACT

A robust method for simultaneous visualization of all four cell cycle phases in living cells is highly desirable. We developed an intensiometric reporter of the transition from S to G2 phase and engineered a far-red fluorescent protein, mMaroon1, to visualize chromatin condensation in mitosis. We combined these new reporters with the previously described Fucci system to create Fucci4, a set of four orthogonal fluorescent indicators that together resolve all cell cycle phases.


Subject(s)
Cell Cycle/physiology , Luminescent Proteins/chemistry , Molecular Imaging/methods , Recombinant Fusion Proteins/chemistry , Time-Lapse Imaging/methods , Animals , Cell Culture Techniques , Chromatin/metabolism , G2 Phase/physiology , HEK293 Cells , HeLa Cells , Humans , Luminescent Proteins/genetics , Mice , Mitosis , Models, Molecular , NIH 3T3 Cells , Recombinant Fusion Proteins/genetics , S Phase/physiology , Red Fluorescent Protein
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